CN114507747B - Tobacco SNP marker developed based on whole genome re-sequencing and KASP technology and application thereof - Google Patents

Tobacco SNP marker developed based on whole genome re-sequencing and KASP technology and application thereof Download PDF

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CN114507747B
CN114507747B CN202111086019.9A CN202111086019A CN114507747B CN 114507747 B CN114507747 B CN 114507747B CN 202111086019 A CN202111086019 A CN 202111086019A CN 114507747 B CN114507747 B CN 114507747B
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余世洲
杨志晓
陈勇
王丰
刘胜杰
雷波
任学良
余婧
赵会纳
张剑锋
金静静
王兵
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Guizhou Institute of Tobacco Science
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Abstract

The invention discloses a tobacco SNP marker developed based on whole genome re-sequencing and KASP technology and application thereof. The method is based on 150 parts of tobacco whole genome re-sequencing data, utilizes a data analysis method of early exploration to mine and obtain more than one million SNP sites, further analyzes the sites, utilizes a KASP technical platform to identify and screen 240 core SNP markers and matched KASP primers, and covers 24 chromosomes/linkage groups of tobacco. The invention provides a set of credible SNP markers which are convenient to be utilized in later period for the research of tobacco biology, and the set of SNP markers and detection primers can be used for the development of a tobacco SNP detection kit, the construction of a tobacco variety DNA fingerprint, the genotyping of tobacco germplasm resources and genetic population, the detection of the background recovery rate of tobacco directionally improved varieties, the detection of the purity/authenticity of tobacco varieties, the auxiliary selection breeding of tobacco molecular markers and the like.

Description

Tobacco SNP marker developed based on whole genome re-sequencing and KASP technology and application thereof
Technical Field
The invention relates to the technical field of biology, in particular to a tobacco SNP marker developed based on whole genome re-sequencing and KASP technology and application thereof.
Background
In the future, china and even the world face various problems such as global climate change, land resource shortage, ecological environment pollution, population quantity surge and the like, a series of challenges are brought to biological breeding, and a traditional breeding technology is urgently combined with a modern molecular biotechnology and an information technology through technological innovation to reform a breeding method so as to meet the requirements of future development on biological excellent varieties.
Currently, with the development of sequencing technologies, reference genomes of main biological species are continuously published, and research projects such as whole genome re-sequencing, simplified genome sequencing, transcriptome sequencing and the like of various species based on sequencing technologies are continuously broken through. The breakthrough of the technologies is fed back to the field of biological breeding, so that the current biological breeding technology gradually gets rid of the past stage of relying on observation of field phenotypic variation, and the stages of time and labor waste and low efficiency are realized by means of knowledge technologies such as statistics, genetics and the like. With the help of the development of various high-throughput sequencing and chip platforms, molecular marker-assisted selective breeding technology is gradually applied to crop breeding, and since the last 80 centuries, the potential of DNA (deoxyribonucleic acid) -based molecular markers in the field of crop commercial breeding is proved, and the molecular markers are widely adopted by a plurality of breeding companies and scientific research units at home and abroad.
According to the state of the art, DNA molecular markers can be divided into three generations. The first generation molecular marker is an RFLP (restriction fragment length polymorphism) marker based on a Southern hybridization technology, the second generation molecular marker is mainly RAPD (random amplification polymorphism), SSR (microsatellite marker), AFLP (amplified fragment length polymorphism), SCAR (characteristic sequence amplification region), STS (specific sequence site), ISSR (internal sequence repeat) and CAPS (restriction enzyme digestion amplification polymorphic sequence) markers which are developed for the core based on a PCR technology, and the third generation molecular marker is mainly SNP (single nucleotide polymorphism). The SNP markers have the advantages of large quantity, uniform distribution in genomes, easy genotyping, suitability for large-scale high-throughput detection and the like, and are used as the most mainstream molecular markers in crop breeding such as corn, wheat, cotton, rice, soybean, rape and the like.
The SNP marker analysis technology can be divided into two categories, SNP marker development and SNP marker detection according to research objects, wherein the SNP marker development mainly comprises searching unknown SNP and analyzing the unknown SNP, and the SNP marker detection comprises detecting SNP genetic diversity of different materials and genotyping.
Tobacco is one of model organisms for researching plants, is a common tool in the field of plant molecular research, is also used as an economic crop to generate great value to agricultural economy, and due to the importance of tobacco in scientific research and industrial economy, large-scale tobacco whole genome sequencing work is carried out in multiple countries such as the United states, japan, china and the like, so that massive tobacco genome sequencing data are accumulated, and a good foundation is laid for development and breeding utilization of tobacco SNP markers. In recent years, some SNP markers have been developed by domestic research units based on means such as EST (expressed sequence tags), GBS (genome reduction methods), RAD (restriction endonuclease site tags), RNAseq (transcriptome sequencing) and the like, but these markers have some defects in terms of genome coverage, are the results of data analysis, lack experimental verification, and provide no detection technical support for the developed SNP markers, so that the developed SNP markers are limited in practical application.
Disclosure of Invention
The invention aims to provide a tobacco SNP marker developed based on whole genome re-sequencing and KASP technology and application thereof.
In a first aspect, the invention claims the use of all or part of the following 240 SNP sites in the tobacco genome in any one of:
(1) Developing a tobacco SNP locus detection kit; the SNP sites referred to herein are all or some of the following 240 SNP sites;
(2) Genotyping a tobacco germplasm resource and/or genetic population;
(3) Constructing a DNA fingerprint of the tobacco germplasm resources/varieties;
(4) And detecting the purity and/or the authenticity of the tobacco variety.
The physical positions of the 240 SNP loci are determined based on the comparison of the whole genome sequence of the tobacco variety Hongda, and the version number of the whole genome sequence of the tobacco variety Hongda is V1.2; the 240 SNP loci are Ts001-Ts240;
ts001 is located on chromosome 1 at position 8208363 with the deoxynucleotide being T or C;
ts002 is located at 33067181 on chromosome 1, and its deoxynucleotide is G or A;
ts003 is located at position 57320321 on chromosome 1, and its deoxynucleotide is G or A;
ts004 is located at position 81583864 of chromosome 1, and the deoxynucleotide of the Ts is A or C;
ts005 is located at 106322334 of chromosome 1, and its deoxynucleotide is G or A;
ts006 is located on chromosome 1 at position 125569732, and its deoxynucleotide is T or C;
ts007 is located at position 146339969 on chromosome 1, and the deoxynucleotide of the Ts is A or G;
ts008 is located at 168939497 th position on chromosome 1, and the deoxynucleotide is A or G;
ts009 is located at position 192328872 on chromosome 1 with deoxynucleotides a or G;
ts010 is located at 218318116 of chromosome 1, and its deoxynucleotide is C or T;
ts011 is located at position 450633 on chromosome 2, and the deoxynucleotide of the Ts011 is C or T;
ts012 is located at position 27809195 on chromosome 2, and its deoxynucleotide is A or T;
ts013 is located at 44225412 on chromosome 2, and its deoxynucleotide is C or T;
ts014 is located at position 62278727 on chromosome 2, and its deoxynucleotide is C or T;
ts015 is located at position 83667001 of chromosome 2, its deoxynucleotide is A or G;
ts016 is located on chromosome 2 at position 113116215, and its deoxynucleotide is G or A;
ts017 is positioned at 137528012 th position on chromosome 2, and the deoxynucleotide is G or A;
ts018 is located on chromosome 2 at position 169132158 with the deoxynucleotide being T or C;
ts019 is located at position 185483126 on chromosome 2 with the deoxynucleotide being G or A;
ts020 is located on chromosome 2 at position 208449625, and its deoxynucleotide is C or T;
ts021 is positioned at 70093 th position on chromosome 3, and the deoxynucleotide is T or C;
ts022 is positioned at the 27066915 th site on the chromosome 3, and the deoxynucleotide of the Ts is A or G;
ts023 is located at position 54763744 on chromosome 3 with the deoxynucleotide G or a;
ts024 is located at position 70866040 on chromosome 3, and its deoxynucleotide is A or G;
ts025 is located at position 91669016 on chromosome 3, and its deoxynucleotide is C or T;
ts026 is located on chromosome 3 at position 116024385, and its deoxynucleotide is G or A;
ts027 is located on chromosome 3 at position 136069307, with the deoxynucleotide being T or G;
ts028 is located on chromosome 3 at position 156967681, and its deoxynucleotide is G or A;
ts029 is located on chromosome 3 at position 173064283, and the deoxynucleotide is A or G;
ts030 is located on chromosome 3 at position 198017426, and its deoxynucleotide is T or C;
ts031 is located on chromosome 4 at position 3899155 with deoxynucleotides G or A;
ts032 is located at position 15517251 on chromosome 4, and the deoxynucleotide is C or T;
ts033 is located at position 26648428 on chromosome 4, and its deoxynucleotide is C or G;
ts034 is located on chromosome 4 at position 46186327 and its deoxynucleotide is C or T;
ts035 is 63336872 on chromosome 4, and its deoxynucleotide is C or G;
ts036 is located at position 83168757 on chromosome 4, and its deoxynucleotide is A or G;
ts037 is located at position 94688224 on chromosome 4, and its deoxynucleotide is C or T;
ts038 is located on chromosome 4 at position 105682078, and its deoxynucleotide is C or T;
ts039 is located at position 124010732 on chromosome 4 with the deoxynucleotide being C or G;
ts040 is located at position 136418400 on chromosome 4, and its deoxynucleotide is C or G;
ts041 is located at position 818540 on chromosome 5, and its deoxynucleotide is G or C;
ts042 is located on chromosome 5 at position 13565159, and the deoxynucleotide is G or T;
ts043 is located on chromosome 5 at position 27317490, and the deoxynucleotide is C or T;
ts044 is located on chromosome 5 at position 42181447, and the deoxynucleotide is G or A;
ts045 is located at 67747378 th position on chromosome 5, and the deoxynucleotide is A or T;
ts046 is located at 80270025 th position on chromosome 5, and the deoxynucleotide is G or T;
ts047 is located on chromosome 5 at position 88119523, and its deoxynucleotide is A or G;
ts048 is located on chromosome 5 at position 98029128, and its deoxynucleotide is T or A;
ts049 is located on chromosome 5 at position 127826992, and its deoxynucleotide is A or T;
ts050 is located at position 133049305 on chromosome 5, and the deoxynucleotide of the Ts is A or T;
ts051 is located at position 13578189 on chromosome 6, and the deoxynucleotide is C or T;
ts052 is located on chromosome 6 at position 25353358 with the deoxynucleotide being G or A;
ts053 is located on chromosome 6 at position 4424521, and its deoxynucleotide is T or C;
ts054 is located at position 58884603 on chromosome 6, and its deoxynucleotide is G or A;
ts055 is located at 72393591 th site on chromosome 6, and the deoxynucleotide is A or G;
ts056 is located at position 84232195 on chromosome 6, and its deoxynucleotide is C or T;
ts057 is located at 95593235 on chromosome 6, and its deoxynucleotide is A or C;
ts058 is located on chromosome 6 at position 115033653, and its deoxynucleotide is T or A;
ts059 is located on chromosome 6 at position 125125703 and its deoxynucleotide is A or G;
ts060 is located on chromosome 6 at position 141918758, and its deoxynucleotide is C or T;
ts061 is located on chromosome 7 at position 414141224 with deoxynucleotides C or T;
ts062 is located at position 14082999 on chromosome 7, and its deoxynucleotide is C or A;
ts063 is located at 22079279 on chromosome 7, and its deoxynucleotide is C or T;
ts064 is located at position 36706020 on chromosome 7, and the deoxynucleotide is G or T;
ts065 is located on chromosome 7 at position 51446290, and its deoxynucleotide is T or C;
ts066 is located at position 64707494 on chromosome 7, and the deoxynucleotide of the Ts is T or A;
ts067 is located at 79594403 th position on chromosome 7, and the deoxynucleotide of the Ts067 is G or A;
ts068 is located at position 100880287 on chromosome 7 with the deoxynucleotide being C or G;
ts069 is located at the 116533979 position on chromosome 7, and the deoxynucleotide is A or C;
ts070 is located at position 134235128 on chromosome 7, and the deoxynucleotide is A or T;
ts071 is located at 3230578 th position on chromosome 8, and its deoxynucleotide is G or A;
ts072 is located on chromosome 8 at position 14779614 with deoxynucleotides C or T;
ts073 is located on chromosome 8 at position 34418092, and its deoxynucleotide is C or A;
ts074 is located on chromosome 8 at position 47428516, and its deoxynucleotide is G or A;
ts075 is located on chromosome 8 at position 64510378, and its deoxynucleotide is G or T;
ts076 is located at position 77171808 on chromosome 8, and its deoxynucleotide is A or C;
ts077 is located on chromosome 8 at position 86364114, and its deoxynucleotide is T or C;
ts078 is located on chromosome 8 at position 103332416, and its deoxynucleotide is A or G;
ts079 is located at 119385289 on chromosome 8, and its deoxynucleotide is C or G;
ts080 is located at 139629563 position on chromosome 8, and the deoxynucleotide is C or T;
ts081 is located at position 10705780 on chromosome 9, and the deoxynucleotide is A or T;
ts082 is located at position 25510126 on chromosome 9, and its deoxynucleotide is A or G;
ts083 is located at position 36791110 on chromosome 9, and its deoxynucleotide is C or T;
ts084 is located on chromosome 9 at position 52279521 with deoxynucleotides C or A;
ts085 is located at position 66389919 on chromosome 9, and its deoxynucleotide is A or G;
ts086 is located on chromosome 9 at position 76596627 with deoxynucleotide T or C;
ts087 is located at position 88782369 on chromosome 9, and its deoxynucleotide is A or G;
ts088 is located at position 101792480 on chromosome 9, and the deoxynucleotide is A or G;
ts089 is located on chromosome 9 at position 114504225, and its deoxynucleotide is A or G;
ts090 is located at position 125921087 on chromosome 9, and the deoxynucleotide is C or T;
ts091 is located on chromosome 10 at position 8333061, and its deoxynucleotide is T or G;
ts092 is located on chromosome 10 at position 19900537 with a deoxynucleotide that is T or C;
ts093 is located on chromosome 10 at position 30838441, and its deoxynucleotide is A or G;
ts094 is located on chromosome 10 at position 48459980 with a deoxynucleotide A or T;
ts095 is located on chromosome 10 at position 60720808, and its deoxynucleotide is A or G;
ts096 is located on chromosome 10 at position 81556459, and its deoxynucleotide is G or C;
ts097 is located on chromosome 10 at position 91772677, and its deoxynucleotide is G or A;
ts098 is located on chromosome 10 at position 104792976 with a deoxynucleotide as G or A;
ts099 is located on chromosome 10 at position 115447057 with a deoxynucleotide G or A;
ts100 is located on chromosome 10 at position 127459715, and the deoxynucleotide of the Ts is A or G;
ts101 is located at 1849343 on chromosome 11, and its deoxynucleotide is C or T;
ts102 is located at position 9349425 on chromosome 11, and its deoxynucleotide is A or G;
ts103 is located at 21455270 th on chromosome 11, and the deoxynucleotide is C or T;
ts104 is positioned at position 26997488 on chromosome 11, and the deoxynucleotide is C or T;
ts105 is located on chromosome 11 at position 39424005, and its deoxynucleotide is T or G;
ts106 is located at the 54261766 position on chromosome 11, and the deoxynucleotide of the Ts is G or T;
ts107 is located on chromosome 11 at position 71261702 with the deoxynucleotide being T or G;
ts108 is located on chromosome 11 at position 89921212, and the deoxynucleotide is T or A;
ts109 is located on chromosome 11 at position 108191371, and its deoxynucleotide is G or A;
ts110 is located on chromosome 11 at position 125710919, and its deoxynucleotide is C or T;
ts111 is located on chromosome 12 at position 6346289, and its deoxynucleotide is G or A;
ts112 is located on chromosome 12 at position 19411089, and its deoxynucleotide is G or C;
ts113 is located at position 35572511 on chromosome 12, and its deoxynucleotide is C or A;
ts114 is located on chromosome 12 at position 50563895, and its deoxynucleotide is C or A;
ts115 is located at position 64428830 on chromosome 12, and the deoxynucleotide of the Ts is A or G;
ts116 is located at the position 72201030 on chromosome 12, and the deoxynucleotide of the Ts is C or T;
ts117 is located on chromosome 12 at position 84715140 with deoxynucleotide A or G;
ts118 is located on chromosome 12 at position 96783131 with the deoxynucleotide being C or T;
ts119 is located on chromosome 12 at position 112836712, and its deoxynucleotide is C or A;
ts120 is located on chromosome 12 at position 120317419, and its deoxynucleotide is C or T;
ts121 is located on chromosome 13 at position 2084818 with the deoxynucleotide being a or G;
ts122 is located on chromosome 13 at position 15919260, and its deoxynucleotide is A or T;
ts123 is located at 29366753 th site on chromosome 13, and the deoxynucleotide is G or T;
ts124 is located on the No.13 chromosome at the 42405167 position, and the deoxynucleotide of the Ts is G or A;
ts125 is located on chromosome 13 at position 50730421 with a deoxynucleotide T or C;
ts126 is located on chromosome 13 at position 60261050 with a deoxynucleotide that is G or a;
ts127 is located on chromosome 13 at position 71762485, and its deoxynucleotide is C or A;
ts128 is located on chromosome 13 at position 86588177, and its deoxynucleotide is C or T;
ts129 is located at position 100565007 on chromosome 13, and the deoxynucleotide of the Ts is A or G;
ts130 is located at position 111060065 on chromosome 13, and the deoxynucleotide is T or G;
ts131 is located on chromosome 14 at position 761393, and its deoxynucleotide is G or A;
ts132 is located at position 10247910 on chromosome 14 with the deoxynucleotide C or T;
ts133 is located at position 25145170 on chromosome 14 with the deoxynucleotide being G or A;
ts134 is located on chromosome 14 at position 33808789, and its deoxynucleotide is A or G;
ts135 is located on chromosome 14 at position 43276550, and its deoxynucleotide is G or A;
ts136 is located on chromosome 14 at position 54717027 with the deoxynucleotide being T or C;
ts137 is located at position 64685001 on chromosome 14, and its deoxynucleotide is G or T;
ts138 is located at position 76697305 on chromosome 14, and the deoxynucleotide of the Ts is C or T;
ts139 is located on chromosome 14 at position 89419411, and its deoxynucleotide is A or T;
ts140 is located on chromosome 14 at position 105075736 with the deoxynucleotide a or G;
ts141 is located on chromosome 15 at position 151458 with the deoxynucleotide being T or C;
ts142 is located at position 9949166 on chromosome 15, and its deoxynucleotide is G or A;
ts143 is located on chromosome 15 at position 20982657, and its deoxynucleotide is A or G;
ts144 is located on chromosome 15 at position 31321347 with the deoxynucleotide being G or T;
ts145 is located on chromosome 15 at position 43232491, and its deoxynucleotide is A or C;
ts146 is located at the 49438846 th position on chromosome 15, and the deoxynucleotide of the Ts is G or C;
ts147 is located at 65072165 position on chromosome 15, and the deoxynucleotide is C or T;
ts148 is located on chromosome 15 at position 76351185, and its deoxynucleotide is A or C;
ts149 is located on chromosome 15 at position 89189533 with the deoxynucleotide being G or A;
ts150 is located on chromosome 15 at position 104194974 with the deoxynucleotide being T or G;
ts151 is located on chromosome 16 at position 1110051, and its deoxynucleotide is T or C;
ts152 is located at 10087681 position on chromosome 16, and the deoxynucleotide of the Ts is C or T;
ts153 is located at 17842746 th on chromosome 16, and the deoxynucleotide is C or T;
ts154 is located at position 26067906 on chromosome 16, and its deoxynucleotide is A or C;
ts155 is located on chromosome 16 at position 34995199, and its deoxynucleotide is G or A;
ts156 is located on chromosome 16 at position 46203805, and its deoxynucleotide is T or C;
ts157 is located on chromosome 16 at position 52542790 with the deoxynucleotide being T or C;
ts158 is located on chromosome 16 at position 62428942, and its deoxynucleotide is C or G;
ts159 is located on chromosome 16 at position 79498876 with the deoxynucleotide being C or T;
ts160 is located on chromosome 16 at position 82884529 with the deoxynucleotide C or a;
ts161 is located on chromosome 17 at position 3824750 with the deoxynucleotide being A or G;
ts162 is located on chromosome 17 at position 13454308 with the deoxynucleotide being C or T;
ts163 is located on chromosome 17 at position 23361027 with the deoxynucleotide being T or C;
ts164 is located on chromosome 17 at position 30960072, and its deoxynucleotide is T or A;
ts165 is located on chromosome 17 at position 3985348, and its deoxynucleotide is G or A;
ts166 is located on chromosome 17 at position 49241377, and its deoxynucleotide is G or A;
ts167 is located on chromosome 17 at position 56588921 with deoxynucleotides a or G;
ts168 is located on chromosome 17 at position 68788574 with the deoxynucleotide being T or a;
ts169 is located on chromosome 17 at position 76261437, and its deoxynucleotide is A or T;
ts170 is located on chromosome 17 at position 86191077 with deoxynucleotides C or T;
ts171 is located at position 10666388 on chromosome 18, and its deoxynucleotide is T or G;
ts172 is located on chromosome 18 at position 23470800 with the deoxynucleotide a or G;
ts173 is located on chromosome 18 at position 30405985 with a deoxynucleotide of C or T;
ts174 is located on chromosome 18 at position 37097793, and its deoxynucleotide is C or A;
ts175 is located on chromosome 18 at position 41051715 with the deoxynucleotide being T or A;
ts176 is located on chromosome 18 at position 49088497 with deoxynucleotide C or A;
ts177 is located on chromosome 18 at position 58652117 with the deoxynucleotide being G or T;
ts178 is located on chromosome 18 at position 69344463 with a deoxynucleotide of T or C;
ts179 is located at position 77733306 on chromosome 18, and its deoxynucleotide is T or A;
ts180 is located on chromosome 18 at position 84321700, and its deoxynucleotide is G or A;
ts181 is located at position 2073784 on chromosome 19 with the deoxynucleotide being T or A;
ts182 is located on chromosome 19 at position 7537621 with the deoxynucleotide being G or a;
ts183 is located on chromosome 19 at position 13153286, and its deoxynucleotide is C or T;
ts184 is located at 17723705 th site on chromosome 19, and its deoxynucleotide is C or G;
ts185 is located on chromosome 19 at position 27550742 with the deoxynucleotide being T or C;
ts186 is located on chromosome 19 at position 32885827 with the deoxynucleotide being a or G;
ts187 is located at position 39636484 on chromosome 19, and its deoxynucleotide is G or A;
ts188 is located on chromosome 19 at position 44936247, and the deoxynucleotide is C or T;
ts189 is located at 57465608 on chromosome 19, and its deoxynucleotide is C or T;
ts190 is located on chromosome 19 at position 74639708, and its deoxynucleotide is C or T;
ts191 is located on chromosome 20 at position 4182297 with the deoxynucleotide being G or A;
ts192 is located at position 13966980 on chromosome 20 with the deoxynucleotide being G or a;
ts193 is located on chromosome 20 at position 19609796 with the deoxynucleotide being G or a;
ts194 is located at position 29237378 on chromosome 20, and the deoxynucleotide is A or G;
ts195 is located on chromosome 20 at position 36180655, and its deoxynucleotide is T or G;
ts196 is located at position 39861635 on chromosome 20, and its deoxynucleotide is G or C;
ts197 is located on chromosome 20 at position 52409493, and its deoxynucleotide is T or A;
ts198 is positioned at the 56616994 th site on the 20 th chromosome, and the deoxynucleotide is C or T;
ts199 is located on chromosome 20 at position 66112600 with the deoxynucleotide being G or T;
ts200 is located on chromosome 20 at position 76769655 with the deoxynucleotide being G or A;
ts201 is located at position 1120311 on chromosome 21, and its deoxynucleotide is A or G;
ts202 is located on chromosome 21 at position 8841119 with the deoxynucleotide being T or G;
ts203 is located at position 15008035 on chromosome 21, and the deoxynucleotide is G or A;
ts204 is located at position 21697573 on chromosome 21, and the deoxynucleotide is T or G;
ts205 is located on chromosome 21 at position 29569502, and its deoxynucleotide is C or T;
ts206 is located at position 35545304 on chromosome 21, and its deoxynucleotide is A or T;
ts207 is located on chromosome 21 at position 43960675, and the deoxynucleotide is C or T;
ts208 is located on chromosome 21 at position 52269430, and the deoxynucleotide is A or G;
ts209 is located on the 60583608 th site on chromosome 21, and the deoxynucleotide of the Ts is A or G;
ts210 is located at 72870875 th position on chromosome 21, and the deoxynucleotide of the Ts is A or C;
ts211 is located on chromosome 22 at position 473170, and its deoxynucleotide is A or G;
ts212 is located on chromosome 22 at position 8509606 with the deoxynucleotide being T or C;
ts213 is located on chromosome 22 at position 12853488, and its deoxynucleotide is T or C;
ts214 is located at 22117353 th position on chromosome 22, and the deoxynucleotide of the Ts is G or A;
ts215 is located at position 26228906 on chromosome 22 with the deoxynucleotide being T or A;
ts216 is located on chromosome 22 at position 33899956, and its deoxynucleotide is C or T;
ts217 is located on chromosome 22 at position 41179872 with the deoxynucleotide being C or a;
ts218 is located on chromosome 22 at position 47452531 with the deoxynucleotide C or G;
ts219 is located on chromosome 22 at position 58059052, and its deoxynucleotide is T or A;
ts220 is located at position 72943832 on chromosome 22, and its deoxynucleotide is A or G;
ts221 is located at position 79892 on chromosome 23 with the deoxynucleotide C or T;
ts222 is located on chromosome 23 at position 5555730, and its deoxynucleotide is C or T;
ts223 is located on chromosome 23 at position 13135791 with deoxynucleotides A or G;
ts224 is located on chromosome 23 at position 24118597 with the deoxynucleotide being A or T;
ts225 is located on chromosome 23 at position 34819441, and the deoxynucleotide is G or A;
ts226 is located on chromosome 23 at position 42069769, and its deoxynucleotide is T or C;
ts227 is located on chromosome 23 at position 46079860, with the deoxynucleotide being G or a;
ts228 is located on chromosome 23 at position 50972058 with the deoxynucleotide being a or G;
ts229 is located on chromosome 23 at position 58370965, with the deoxynucleotide being C or T;
ts230 is located on chromosome 23 at position 6624863, and its deoxynucleotide is A or T;
ts231 is located on chromosome 24 at position 5769442, and its deoxynucleotide is C or T;
ts232 is located on chromosome 24 at position 16375603, and its deoxynucleotide is T or C;
ts233 is located on chromosome 24 at position 19508580, and its deoxynucleotide is G or A;
ts234 is located on chromosome 24 at position 20893673, and its deoxynucleotide is T or G;
ts235 is located on chromosome 24 at position 27208986, and its deoxynucleotide is T or C;
ts236 is located on chromosome 24 at position 34358980, and its deoxynucleotide is C or T;
ts237 is located at position 44075839 on chromosome 24, and its deoxynucleotide is T or G;
ts238 is located on chromosome 24 at position 49853976, and its deoxynucleotide is A or G;
ts239 is located at position 58945156 on chromosome 24, and the deoxynucleotide of the Ts239 is T or G;
ts240 is located on chromosome 24 at position 63099098 with a deoxynucleotide as C or G.
Further, all or part of the following 240 SNP sites in the tobacco genome are preferably 48 SNP site combinations as follows: ts003, ts007, ts013, ts019, ts028, ts029, ts033, ts039, ts041, ts048, ts055, ts060, ts062, ts070, ts074, ts079, ts082, ts089, ts093, ts098, ts106, ts108, ts113, ts120, ts122, ts129, ts132, ts138, ts141, ts148, ts152, ts159, ts164, ts169, ts172, ts179, ts183, ts186, ts192, ts194, ts201, ts207, ts212, ts218, ts222, ts230, ts232 and Ts236.
Further, the single nucleotide polymorphisms of the 240 SNP sites and the upstream and downstream 50bp sequences are shown in Table 3.
In a second aspect, the invention claims sets of KASP primers.
The set of KASP primers claimed in the present invention are KASP primers for detecting all or part of the 240 SNP sites described in the first aspect above.
Further, the set of KASP primers is preferably a set of KASP primers for detecting the following 48 SNP sites: ts003, ts007, ts013, ts019, ts028, ts029, ts033, ts039, ts041, ts048, ts055, ts060, ts062, ts070, ts074, ts079, ts082, ts089, ts093, ts098, ts106, ts108, ts113, ts120, ts122, ts129, ts132, ts138, ts141, ts148, ts152, ts159, ts164, ts169, ts172, ts179, ts183, ts186, ts192, ts194, ts201, ts207, ts212, ts218, ts222, ts230, ts232 and Ts236.
The KASP primers corresponding to each SNP site to be detected respectively comprise two forward primers and one reverse primer; wherein two forward primers are respectively marked as a forward primer 1 and a forward primer 2; the 5 'end of the forward primer 1 is connected with one fluorescent label sequence, and the 5' end of the forward primer 2 is connected with the other fluorescent label sequence.
The nucleotide sequence of a forward primer 1 for detecting Ts001 is shown as SEQ ID No.1, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.241, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 481;
the nucleotide sequence of the forward primer 1 for detecting Ts002 is shown as SEQ ID No.2, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.242, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 482;
the nucleotide sequence of a forward primer 1 for detecting Ts003 is shown as SEQ ID No.3, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.243, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 483;
the nucleotide sequence of the forward primer 1 for detecting Ts004 is shown as SEQ ID No.4, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.244, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 484;
the nucleotide sequence of the forward primer 1 for detecting Ts005 is shown as SEQ ID No.5, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.245, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 485;
the nucleotide sequence of a forward primer 1 for detecting Ts006 is shown as SEQ ID No.6, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.246, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 486;
the nucleotide sequence of a forward primer 1 for detecting Ts007 is shown as SEQ ID No.7, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.247, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 487;
the nucleotide sequence of a forward primer 1 for detecting Ts008 is shown as SEQ ID No.8, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.248, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 488;
the nucleotide sequence of the forward primer 1 for detecting Ts009 is shown as SEQ ID No.9, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.249, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 489;
the nucleotide sequence of the forward primer 1 for detecting Ts010 is shown as SEQ ID No.10, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.250, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 490;
the nucleotide sequence of a forward primer 1 for detecting Ts011 is shown in SEQ ID No.11, the nucleotide sequence of a forward primer 2 is shown in SEQ ID No.251, and the nucleotide sequence of a reverse primer is shown in SEQ ID No. 491;
the nucleotide sequence of a forward primer 1 for detecting Ts012 is shown as SEQ ID No.12, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.252, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 492;
the nucleotide sequence of a forward primer 1 for detecting Ts013 is shown in SEQ ID No.13, the nucleotide sequence of a forward primer 2 is shown in SEQ ID No.253, and the nucleotide sequence of a reverse primer is shown in SEQ ID No. 493;
the nucleotide sequence of a forward primer 1 for detecting Ts014 is shown as SEQ ID No.14, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.254, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 494;
the nucleotide sequence of a forward primer 1 for detecting Ts015 is shown as SEQ ID No.15, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.255, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 495;
the nucleotide sequence of a forward primer 1 for detecting Ts016 is shown as SEQ ID No.16, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.256, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 496;
the nucleotide sequence of a forward primer 1 for detecting Ts017 is shown as SEQ ID No.17, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.257, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 497;
the nucleotide sequence of a forward primer 1 for detecting Ts018 is shown as SEQ ID No.18, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.258, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 498;
the nucleotide sequence of the forward primer 1 for detecting Ts019 is shown as SEQ ID No.19, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.259, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 499;
the nucleotide sequence of the forward primer 1 for detecting Ts020 is shown in SEQ ID No.20, the nucleotide sequence of the forward primer 2 is shown in SEQ ID No.260, and the nucleotide sequence of the reverse primer is shown in SEQ ID No. 500;
the nucleotide sequence of a forward primer 1 for detecting Ts021 is shown as SEQ ID No.21, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.261, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 501;
the nucleotide sequence of the forward primer 1 for detecting Ts022 is shown as SEQ ID No.22, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.262, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 502;
the nucleotide sequence of the forward primer 1 for detecting Ts023 is shown as SEQ ID No.23, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.263, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 503;
the nucleotide sequence of a forward primer 1 for detecting Ts024 is shown as SEQ ID No.24, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.264, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 504;
the nucleotide sequence of the forward primer 1 for detecting Ts025 is shown as SEQ ID No.25, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.265, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 505;
the nucleotide sequence of a forward primer 1 for detecting Ts026 is shown as SEQ ID No.26, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.266, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 506;
the nucleotide sequence of the forward primer 1 for detecting Ts027 is shown in SEQ ID No.27, the nucleotide sequence of the forward primer 2 is shown in SEQ ID No.267, and the nucleotide sequence of the reverse primer is shown in SEQ ID No. 507;
the nucleotide sequence of a forward primer 1 for detecting Ts028 is shown as SEQ ID No.28, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.268, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 508;
the nucleotide sequence of a forward primer 1 for detecting Ts029 is shown as SEQ ID No.29, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.269, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 509;
the nucleotide sequence of a forward primer 1 for detecting Ts030 is shown as SEQ ID No.30, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.270, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 510;
the nucleotide sequence of the forward primer 1 for detecting Ts031 is shown as SEQ ID No.31, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.271, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 511;
the nucleotide sequence of the forward primer 1 for detecting Ts032 is shown as SEQ ID No.32, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.272, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 512;
the nucleotide sequence of a forward primer 1 for detecting Ts033 is shown as SEQ ID No.33, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.273, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 513;
the nucleotide sequence of a forward primer 1 for detecting Ts034 is shown as SEQ ID No.34, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.274, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 514;
the nucleotide sequence of a forward primer 1 for detecting Ts035 is shown as SEQ ID No.35, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.275, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 515;
the nucleotide sequence of the forward primer 1 for detecting Ts036 is shown as SEQ ID No.36, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.276, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 516;
the nucleotide sequence of the forward primer 1 for detecting Ts037 is shown as SEQ ID No.37, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.277, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 517;
the nucleotide sequence of a forward primer 1 for detecting Ts038 is shown as SEQ ID No.38, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.278, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 518;
the nucleotide sequence of a forward primer 1 for detecting Ts039 is shown as SEQ ID No.39, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.279, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 519;
the nucleotide sequence of the forward primer 1 for detecting Ts040 is shown as SEQ ID No.40, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.280, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 520;
the nucleotide sequence of the forward primer 1 for detecting Ts041 is shown as SEQ ID No.41, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.281, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 521;
the nucleotide sequence of a forward primer 1 for detecting Ts042 is shown as SEQ ID No.42, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.282, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 522;
the nucleotide sequence of the forward primer 1 for detecting Ts043 is shown as SEQ ID No.43, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.283, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 523;
the nucleotide sequence of a forward primer 1 for detecting Ts044 is shown as SEQ ID No.44, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.284, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 524;
the nucleotide sequence of the forward primer 1 for detecting Ts045 is shown as SEQ ID No.45, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.285, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 525;
the nucleotide sequence of the forward primer 1 for detecting Ts046 is shown as SEQ ID No.46, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.286, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 526;
the nucleotide sequence of the forward primer 1 for detecting Ts047 is shown as SEQ ID No.47, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.287, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 527;
the nucleotide sequence of the forward primer 1 for detecting Ts048 is shown as SEQ ID No.48, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.288, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 528;
the nucleotide sequence of a forward primer 1 for detecting Ts049 is shown as SEQ ID No.49, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.289, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 529;
the nucleotide sequence of a forward primer 1 for detecting Ts050 is shown as SEQ ID No.50, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.290, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 530;
the nucleotide sequence of a forward primer 1 for detecting Ts051 is shown as SEQ ID No.51, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.291, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 531;
the nucleotide sequence of the forward primer 1 for detecting Ts052 is shown as SEQ ID No.52, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.292, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 532;
the nucleotide sequence of a forward primer 1 for detecting Ts053 is shown as SEQ ID No.53, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.293, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 533;
the nucleotide sequence of the forward primer 1 for detecting Ts054 is shown as SEQ ID No.54, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.294, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 534;
the nucleotide sequence of the forward primer 1 for detecting Ts055 is shown as SEQ ID No.55, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.295, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 535;
the nucleotide sequence of the forward primer 1 for detecting Ts056 is shown as SEQ ID No.56, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.296, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 536;
the nucleotide sequence of the forward primer 1 for detecting Ts057 is shown as SEQ ID No.57, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.297, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 537;
the nucleotide sequence of the forward primer 1 for detecting Ts058 is shown as SEQ ID No.58, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.298, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 538;
the nucleotide sequence of the forward primer 1 for detecting Ts059 is shown as SEQ ID No.59, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.299, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 539;
the nucleotide sequence of a forward primer 1 for detecting Ts060 is shown in SEQ ID No.60, the nucleotide sequence of a forward primer 2 is shown in SEQ ID No.300, and the nucleotide sequence of a reverse primer is shown in SEQ ID No. 540;
the nucleotide sequence of a forward primer 1 for detecting Ts061 is shown as SEQ ID No.61, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.301, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 541;
the nucleotide sequence of a forward primer 1 for detecting Ts062 is shown as SEQ ID No.62, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.302, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 542;
the nucleotide sequence of a forward primer 1 for detecting Ts063 is shown as SEQ ID No.63, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.303, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 543;
the nucleotide sequence of a forward primer 1 for detecting Ts064 is shown as SEQ ID No.64, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.304, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 544;
the nucleotide sequence of a forward primer 1 for detecting Ts065 is shown as SEQ ID No.65, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.305, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 545;
the nucleotide sequence of the forward primer 1 for detecting Ts066 is shown as SEQ ID No.66, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.306, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 546;
the nucleotide sequence of a forward primer 1 for detecting Ts067 is shown as SEQ ID No.67, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.307, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 547;
the nucleotide sequence of a forward primer 1 for detecting Ts068 is shown as SEQ ID No.68, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.308, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 548;
the nucleotide sequence of a forward primer 1 for detecting Ts069 is shown as SEQ ID No.69, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.309, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 549;
the nucleotide sequence of a forward primer 1 for detecting Ts070 is shown as SEQ ID No.70, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.310, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 550;
the nucleotide sequence of a forward primer 1 for detecting Ts071 is shown as SEQ ID No.71, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.311, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 551;
the nucleotide sequence of a forward primer 1 for detecting Ts072 is shown as SEQ ID No.72, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.312, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 552;
the nucleotide sequence of a forward primer 1 for detecting Ts073 is shown as SEQ ID No.73, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.313, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 553;
the nucleotide sequence of the forward primer 1 for detecting Ts074 is shown as SEQ ID No.74, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.314, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 554;
the nucleotide sequence of the forward primer 1 for detecting Ts075 is shown as SEQ ID No.75, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.315, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 555;
the nucleotide sequence of the forward primer 1 for detecting Ts076 is shown as SEQ ID No.76, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.316, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 556;
the nucleotide sequence of a forward primer 1 for detecting Ts077 is shown as SEQ ID No.77, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.317, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 557;
the nucleotide sequence of the forward primer 1 for detecting Ts078 is shown as SEQ ID No.78, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.318, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 558;
the nucleotide sequence of the forward primer 1 for detecting Ts079 is shown as SEQ ID No.79, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.319, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 559;
the nucleotide sequence of a forward primer 1 for detecting Ts080 is shown as SEQ ID No.80, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.320, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 560;
the nucleotide sequence of a forward primer 1 for detecting Ts081 is shown as SEQ ID No.81, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.321, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 561;
the nucleotide sequence of the forward primer 1 for detecting Ts082 is shown as SEQ ID No.82, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.322, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 562;
the nucleotide sequence of a forward primer 1 for detecting Ts083 is shown as SEQ ID No.83, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.323, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 563;
the nucleotide sequence of the forward primer 1 for detecting Ts084 is shown as SEQ ID No.84, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.324, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 564;
the nucleotide sequence of the forward primer 1 for detecting Ts085 is shown as SEQ ID No.85, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.325, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 565;
the nucleotide sequence of the forward primer 1 for detecting Ts086 is shown as SEQ ID No.86, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.326, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 566;
the nucleotide sequence of the forward primer 1 for detecting Ts087 is shown as SEQ ID No.87, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.327, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 567;
the nucleotide sequence of a forward primer 1 for detecting Ts088 is shown as SEQ ID No.88, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.328, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 568;
the nucleotide sequence of a forward primer 1 for detecting Ts089 is shown as SEQ ID No.89, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.329, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 569;
the nucleotide sequence of the forward primer 1 for detecting Ts090 is shown as SEQ ID No.90, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.330, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 570;
the nucleotide sequence of the forward primer 1 for detecting Ts091 is shown as SEQ ID No.91, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.331, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 571;
the nucleotide sequence of the forward primer 1 for detecting Ts092 is shown as SEQ ID No.92, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.332, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 572;
the nucleotide sequence of the forward primer 1 for detecting Ts093 is shown as SEQ ID No.93, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.333, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 573;
the nucleotide sequence of the forward primer 1 for detecting Ts094 is shown as SEQ ID No.94, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.334, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 574;
the nucleotide sequence of the forward primer 1 for detecting Ts095 is shown as SEQ ID No.95, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.335, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 575;
the nucleotide sequence of the forward primer 1 for detecting Ts096 is shown as SEQ ID No.96, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.336, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 576;
the nucleotide sequence of the forward primer 1 for detecting Ts097 is shown as SEQ ID No.97, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.337, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 577;
the nucleotide sequence of the forward primer 1 for detecting Ts098 is shown as SEQ ID No.98, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.338, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 578;
the nucleotide sequence of the forward primer 1 for detecting Ts099 is shown as SEQ ID No.99, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.339, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 579;
the nucleotide sequence of a forward primer 1 for detecting Ts100 is shown as SEQ ID No.100, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.340, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 580;
the nucleotide sequence of a forward primer 1 for detecting Ts101 is shown as SEQ ID No.101, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.341, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 581;
the nucleotide sequence of a forward primer 1 for detecting Ts102 is shown as SEQ ID No.102, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.342, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 582;
the nucleotide sequence of the forward primer 1 for detecting Ts103 is shown as SEQ ID No.103, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.343, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 583;
the nucleotide sequence of the forward primer 1 for detecting Ts104 is shown as SEQ ID No.104, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.344, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 584;
the nucleotide sequence of the forward primer 1 for detecting Ts105 is shown as SEQ ID No.105, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.345, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 585;
the nucleotide sequence of a forward primer 1 for detecting Ts106 is shown as SEQ ID No.106, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.346, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 586;
the nucleotide sequence of a forward primer 1 for detecting Ts107 is shown as SEQ ID No.107, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.347, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 587;
the nucleotide sequence of a forward primer 1 for detecting Ts108 is shown as SEQ ID No.108, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.348, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 588;
the nucleotide sequence of a forward primer 1 for detecting Ts109 is shown as SEQ ID No.109, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.349, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 589;
the nucleotide sequence of a forward primer 1 for detecting Ts110 is shown as SEQ ID No.110, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.350, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 590;
the nucleotide sequence of a forward primer 1 for detecting Ts111 is shown as SEQ ID No.111, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.351, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 591;
the nucleotide sequence of the forward primer 1 for detecting Ts112 is shown as SEQ ID No.112, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.352, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 592;
the nucleotide sequence of a forward primer 1 for detecting Ts113 is shown as SEQ ID No.113, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.353, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 593;
the nucleotide sequence of a forward primer 1 for detecting Ts114 is shown as SEQ ID No.114, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.354, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 594;
the nucleotide sequence of the forward primer 1 for detecting Ts115 is shown as SEQ ID No.115, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.355, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 595;
the nucleotide sequence of the forward primer 1 for detecting Ts116 is shown as SEQ ID No.116, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.356, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 596;
the nucleotide sequence of a forward primer 1 for detecting Ts117 is shown as SEQ ID No.117, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.357, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 597;
the nucleotide sequence of a forward primer 1 for detecting Ts118 is shown as SEQ ID No.118, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.358, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 598;
the nucleotide sequence of a forward primer 1 for detecting Ts119 is shown as SEQ ID No.119, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.359, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 599;
the nucleotide sequence of a forward primer 1 for detecting Ts120 is shown as SEQ ID No.120, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.360, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 600;
the nucleotide sequence of a forward primer 1 for detecting Ts121 is shown as SEQ ID No.121, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.361, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 601;
the nucleotide sequence of the forward primer 1 for detecting Ts122 is shown as SEQ ID No.122, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.362, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 602;
the nucleotide sequence of the forward primer 1 for detecting Ts123 is shown as SEQ ID No.123, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.363, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 603;
the nucleotide sequence of a forward primer 1 for detecting Ts124 is shown as SEQ ID No.124, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.364, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 604;
the nucleotide sequence of the forward primer 1 for detecting Ts125 is shown as SEQ ID No.125, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.365, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 605;
the nucleotide sequence of the forward primer 1 for detecting Ts126 is shown as SEQ ID No.126, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.366, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 606;
the nucleotide sequence of a forward primer 1 for detecting Ts127 is shown as SEQ ID No.127, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.367, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 607;
the nucleotide sequence of a forward primer 1 for detecting Ts128 is shown as SEQ ID No.128, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.368, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 608;
the nucleotide sequence of the forward primer 1 for detecting Ts129 is shown as SEQ ID No.129, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.369, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 609;
the nucleotide sequence of a forward primer 1 for detecting Ts130 is shown as SEQ ID No.130, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.370, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 610;
the nucleotide sequence of the forward primer 1 for detecting Ts131 is shown as SEQ ID No.131, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.371, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 611;
the nucleotide sequence of the forward primer 1 for detecting Ts132 is shown as SEQ ID No.132, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.372, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 612;
the nucleotide sequence of the forward primer 1 for detecting Ts133 is shown as SEQ ID No.133, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.373, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 613;
the nucleotide sequence of the forward primer 1 for detecting Ts134 is shown as SEQ ID No.134, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.374, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 614;
the nucleotide sequence of a forward primer 1 for detecting Ts135 is shown as SEQ ID No.135, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.375, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 615;
the nucleotide sequence of a forward primer 1 for detecting Ts136 is shown as SEQ ID No.136, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.376, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 616;
the nucleotide sequence of a forward primer 1 for detecting Ts137 is shown as SEQ ID No.137, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.377, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 617;
the nucleotide sequence of a forward primer 1 for detecting Ts138 is shown as SEQ ID No.138, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.378, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 618;
the nucleotide sequence of a forward primer 1 for detecting Ts139 is shown as SEQ ID No.139, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.379, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 619;
the nucleotide sequence of a forward primer 1 for detecting Ts140 is shown as SEQ ID No.140, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.380, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 620;
the nucleotide sequence of a forward primer 1 for detecting Ts141 is shown as SEQ ID No.141, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.381, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 621;
the nucleotide sequence of a forward primer 1 for detecting Ts142 is shown as SEQ ID No.142, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.382, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 622;
the nucleotide sequence of a forward primer 1 for detecting Ts143 is shown as SEQ ID No.143, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.383, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 623;
the nucleotide sequence of a forward primer 1 for detecting Ts144 is shown as SEQ ID No.144, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.384, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 624;
the nucleotide sequence of a forward primer 1 for detecting Ts145 is shown as SEQ ID No.145, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.385, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 625;
the nucleotide sequence of a forward primer 1 for detecting Ts146 is shown as SEQ ID No.146, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.386, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 626;
the nucleotide sequence of the forward primer 1 for detecting Ts147 is shown as SEQ ID No.147, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.387, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 627;
the nucleotide sequence of the forward primer 1 for detecting Ts148 is shown as SEQ ID No.148, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.388, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 628;
the nucleotide sequence of a forward primer 1 for detecting Ts149 is shown as SEQ ID No.149, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.389, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 629;
the nucleotide sequence of a forward primer 1 for detecting Ts150 is shown as SEQ ID No.150, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.390, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 630;
the nucleotide sequence of a forward primer 1 for detecting Ts151 is shown as SEQ ID No.151, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.391, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 631;
the nucleotide sequence of a forward primer 1 for detecting Ts152 is shown as SEQ ID No.152, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.392, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 632;
the nucleotide sequence of the forward primer 1 for detecting Ts153 is shown as SEQ ID No.153, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.393, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 633;
the nucleotide sequence of a forward primer 1 for detecting Ts154 is shown as SEQ ID No.154, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.394, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 634;
the nucleotide sequence of the forward primer 1 for detecting Ts155 is shown as SEQ ID No.155, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.395, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 635;
the nucleotide sequence of a forward primer 1 for detecting Ts156 is shown as SEQ ID No.156, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.396, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 636;
the nucleotide sequence of a forward primer 1 for detecting Ts157 is shown as SEQ ID No.157, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.397, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 637;
the nucleotide sequence of a forward primer 1 for detecting Ts158 is shown as SEQ ID No.158, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.398, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 638;
the nucleotide sequence of the forward primer 1 for detecting Ts159 is shown as SEQ ID No.159, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.399, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 639;
the nucleotide sequence of the forward primer 1 for detecting Ts160 is shown as SEQ ID No.160, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.400, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 640;
the nucleotide sequence of a forward primer 1 for detecting Ts161 is shown as SEQ ID No.161, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.401, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 641;
the nucleotide sequence of the forward primer 1 for detecting Ts162 is shown as SEQ ID No.162, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.402, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 642;
the nucleotide sequence of the forward primer 1 for detecting Ts163 is shown as SEQ ID No.163, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.403, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 643;
the nucleotide sequence of a forward primer 1 for detecting Ts164 is shown as SEQ ID No.164, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.404, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 644;
the nucleotide sequence of the forward primer 1 for detecting Ts165 is shown as SEQ ID No.165, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.405, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 645;
the nucleotide sequence of a forward primer 1 for detecting Ts166 is shown as SEQ ID No.166, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.406, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 646;
the nucleotide sequence of the forward primer 1 for detecting Ts167 is shown as SEQ ID No.167, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.407, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 647;
the nucleotide sequence of a forward primer 1 for detecting Ts168 is shown as SEQ ID No.168, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.408, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 648;
the nucleotide sequence of the forward primer 1 for detecting Ts169 is shown as SEQ ID No.169, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.409, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 649;
the nucleotide sequence of the forward primer 1 for detecting Ts170 is shown as SEQ ID No.170, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.410, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 650;
the nucleotide sequence of the forward primer 1 for detecting Ts171 is shown as SEQ ID No.171, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.411, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 651;
the nucleotide sequence of the forward primer 1 for detecting Ts172 is shown as SEQ ID No.172, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.412, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 652;
the nucleotide sequence of a forward primer 1 for detecting Ts173 is shown as SEQ ID No.173, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.413, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 653;
the nucleotide sequence of a forward primer 1 for detecting Ts174 is shown as SEQ ID No.174, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.414, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 654;
the nucleotide sequence of a forward primer 1 for detecting Ts175 is shown as SEQ ID No.175, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.415, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 655;
the nucleotide sequence of the forward primer 1 for detecting Ts176 is shown as SEQ ID No.176, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.416, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 656;
the nucleotide sequence of the forward primer 1 for detecting Ts177 is shown as SEQ ID No.177, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.417, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 657;
the nucleotide sequence of a forward primer 1 for detecting Ts178 is shown as SEQ ID No.178, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.418, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 658;
the nucleotide sequence of the forward primer 1 for detecting Ts179 is shown as SEQ ID No.179, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.419, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 659;
the nucleotide sequence of the forward primer 1 for detecting Ts180 is shown as SEQ ID No.180, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.420, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 660;
the nucleotide sequence of a forward primer 1 for detecting Ts181 is shown as SEQ ID No.181, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.421, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 661;
the nucleotide sequence of a forward primer 1 for detecting Ts182 is shown as SEQ ID No.182, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.422, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 662;
the nucleotide sequence of a forward primer 1 for detecting Ts183 is shown as SEQ ID No.183, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.423, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 663;
the nucleotide sequence of the forward primer 1 for detecting Ts184 is shown as SEQ ID No.184, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.424, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 664;
the nucleotide sequence of a forward primer 1 for detecting Ts185 is shown as SEQ ID No.185, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.425, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 665;
the nucleotide sequence of a forward primer 1 for detecting Ts186 is shown as SEQ ID No.186, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.426, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 666;
the nucleotide sequence of a forward primer 1 for detecting Ts187 is shown as SEQ ID No.187, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.427, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 667;
the nucleotide sequence of the forward primer 1 for detecting Ts188 is shown as SEQ ID No.188, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.428, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 668;
the nucleotide sequence of a forward primer 1 for detecting Ts189 is shown as SEQ ID No.189, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.429, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 669;
the nucleotide sequence of a forward primer 1 for detecting Ts190 is shown as SEQ ID No.190, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.430, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 670;
the nucleotide sequence of a forward primer 1 for detecting Ts191 is shown as SEQ ID No.191, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.431, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 671;
the nucleotide sequence of a forward primer 1 for detecting Ts192 is shown as SEQ ID No.192, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.432, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 672;
the nucleotide sequence of the forward primer 1 for detecting Ts193 is shown as SEQ ID No.193, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.433, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 673;
the nucleotide sequence of a forward primer 1 for detecting Ts194 is shown as SEQ ID No.194, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.434, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 674;
the nucleotide sequence of a forward primer 1 for detecting Ts195 is shown as SEQ ID No.195, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.435, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 675;
the nucleotide sequence of the forward primer 1 for detecting Ts196 is shown as SEQ ID No.196, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.436, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 676;
the nucleotide sequence of a forward primer 1 for detecting Ts197 is shown as SEQ ID No.197, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.437, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 677;
the nucleotide sequence of the forward primer 1 for detecting Ts198 is shown as SEQ ID No.198, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.438, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 678;
the nucleotide sequence of a forward primer 1 for detecting Ts199 is shown as SEQ ID No.199, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.439, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 679;
the nucleotide sequence of a forward primer 1 for detecting Ts200 is shown as SEQ ID No.200, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.440, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 680;
the nucleotide sequence of a forward primer 1 for detecting Ts201 is shown as SEQ ID No.201, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.441, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 681;
the nucleotide sequence of a forward primer 1 for detecting Ts202 is shown as SEQ ID No.202, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.442, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 682;
the nucleotide sequence of a forward primer 1 for detecting Ts203 is shown as SEQ ID No.203, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.443, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 683;
the nucleotide sequence of a forward primer 1 for detecting Ts204 is shown as SEQ ID No.204, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.444, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 684;
the nucleotide sequence of a forward primer 1 for detecting Ts205 is shown as SEQ ID No.205, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.445, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 685;
the nucleotide sequence of a forward primer 1 for detecting Ts206 is shown as SEQ ID No.206, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.446, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 686;
the nucleotide sequence of a forward primer 1 for detecting Ts207 is shown as SEQ ID No.207, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.447, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 687;
the nucleotide sequence of the forward primer 1 for detecting Ts208 is shown as SEQ ID No.208, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.448, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 688;
the nucleotide sequence of a forward primer 1 for detecting Ts209 is shown as SEQ ID No.209, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.449, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 689;
the nucleotide sequence of a forward primer 1 for detecting Ts210 is shown as SEQ ID No.210, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.450, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 690;
the nucleotide sequence of a forward primer 1 for detecting Ts211 is shown as SEQ ID No.211, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.451, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 691;
the nucleotide sequence of the forward primer 1 for detecting Ts212 is shown as SEQ ID No.212, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.452, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 692;
the nucleotide sequence of a forward primer 1 for detecting Ts213 is shown as SEQ ID No.213, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.453, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 693;
the nucleotide sequence of a forward primer 1 for detecting Ts214 is shown as SEQ ID No.214, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.454, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 694;
the nucleotide sequence of a forward primer 1 for detecting Ts215 is shown as SEQ ID No.215, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.455, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 695;
the nucleotide sequence of a forward primer 1 for detecting Ts216 is shown as SEQ ID No.216, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.456, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 696;
the nucleotide sequence of a forward primer 1 for detecting Ts217 is shown as SEQ ID No.217, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.457, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 697;
the nucleotide sequence of a forward primer 1 for detecting Ts218 is shown as SEQ ID No.218, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.458, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 698;
the nucleotide sequence of a forward primer 1 for detecting Ts219 is shown as SEQ ID No.219, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.459, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 699;
the nucleotide sequence of the forward primer 1 for detecting Ts220 is shown as SEQ ID No.220, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.460, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 700;
the nucleotide sequence of the forward primer 1 for detecting Ts221 is shown as SEQ ID No.221, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.461, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 701;
the nucleotide sequence of the forward primer 1 for detecting Ts222 is shown as SEQ ID No.222, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.462, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 702;
the nucleotide sequence of a forward primer 1 for detecting Ts223 is shown as SEQ ID No.223, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.463, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 703;
the nucleotide sequence of a forward primer 1 for detecting Ts224 is shown as SEQ ID No.224, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.464, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 704;
the nucleotide sequence of a forward primer 1 for detecting Ts225 is shown as SEQ ID No.225, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.465, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 705;
the nucleotide sequence of a forward primer 1 for detecting Ts226 is shown as SEQ ID No.226, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.466, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 706;
the nucleotide sequence of the forward primer 1 for detecting Ts227 is shown as SEQ ID No.227, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.467, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 707;
the nucleotide sequence of a forward primer 1 for detecting Ts228 is shown as SEQ ID No.228, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.468, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 708;
the nucleotide sequence of the forward primer 1 for detecting Ts229 is shown as SEQ ID No.229, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.469, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 709;
the nucleotide sequence of the forward primer 1 for detecting Ts230 is shown as SEQ ID No.230, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.470, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 710;
the nucleotide sequence of the forward primer 1 for detecting Ts231 is shown as SEQ ID No.231, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.471, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 711;
the nucleotide sequence of the forward primer 1 for detecting Ts232 is shown as SEQ ID No.232, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.472, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 712;
the nucleotide sequence of a forward primer 1 for detecting Ts233 is shown as SEQ ID No.233, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.473, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 713;
the nucleotide sequence of a forward primer 1 for detecting Ts234 is shown as SEQ ID No.234, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.474, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 714;
the nucleotide sequence of a forward primer 1 for detecting Ts235 is shown as SEQ ID No.235, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.475, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 715;
the nucleotide sequence of a forward primer 1 for detecting Ts236 is shown as SEQ ID No.236, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.476, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 716;
the nucleotide sequence of a forward primer 1 for detecting Ts237 is shown in SEQ ID No.237, the nucleotide sequence of a forward primer 2 is shown in SEQ ID No.477, and the nucleotide sequence of a reverse primer is shown in SEQ ID No. 717;
the nucleotide sequence of a forward primer 1 for detecting Ts238 is shown as SEQ ID No.238, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.478, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 718;
the nucleotide sequence of a forward primer 1 for detecting Ts239 is shown as SEQ ID No.239, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.479, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 719;
the nucleotide sequence of the forward primer 1 for detecting Ts240 is shown as SEQ ID No.240, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.480, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 720.
Further, the fluorescent label sequence connected with the 5' end of the forward primer 1 is an FAM fluorescent label sequence; the fluorescent tag sequence connected with the 5' end of the forward primer 2 is a HEX fluorescent tag sequence.
Furthermore, the FAM fluorescent label sequence is shown as SEQ ID No. 721; the HEX fluorescent label sequence is shown as SEQ ID No. 722.
In the present invention, the set of KASP primers is designated as TaSNP KASP Primermix.
In a third aspect, the invention claims a kit comprising the set of KASP primers tasp KASP primemix as described previously in the second aspect. Further, the reagent or kit may further comprise KASP 2 × Mastermix, mgCl 2 And ddH 2 O。
The KASP 2 x Mastermix in the reagent or the kit contains a fluorescent probe A, a fluorescent probe B, a quenching probe A and a quenching probe B.
The nucleotide sequence of the fluorescent probe A is consistent with the nucleotide sequence of the fluorescent label sequence connected with the 5 'end of the forward primer 1, and the 5' end is connected with a fluorescent group A; the nucleotide sequence of the quenching probe A is reversely complementary with the nucleotide sequence of the label sequence A, and a quenching group is connected to the 3' terminal.
The nucleotide sequence of the fluorescent probe B is consistent with the nucleotide sequence of the fluorescent label sequence connected with the 5 'end of the forward primer 2, and the 5' end is connected with a fluorescent group B; the nucleotide sequence of the quenching probe B is reversely complementary with the nucleotide sequence of the label sequence B, and the 3' terminal is connected with a quenching group.
In a specific embodiment of the invention, the fluorophore a is FAM; the fluorophore B is HXE; the quenching group is BHQ.
In a fourth aspect, the invention claims the use of a set of KASP primers as described in the second aspect above or a kit as described in the third aspect above in any one of:
(1) Developing a tobacco SNP locus detection kit; the SNP sites referred to herein are all or part of the following 240 SNP sites;
(2) Genotyping a tobacco germplasm resource and/or genetic population;
(3) Constructing a DNA fingerprint of tobacco germplasm resources/varieties;
(4) And detecting the purity and/or the authenticity of the tobacco variety.
In a fifth aspect, the invention claims any of the following methods:
the method A comprises the following steps: a method of genotyping a tobacco germplasm resource and/or genetic population comprising the steps of:
(a1) Respectively extracting genome DNA of all tobacco samples in the tested tobacco germplasm resources and/or genetic groups;
(a2) Sequentially preparing a PCR reaction system for the DNA of a tobacco sample to be tested according to the 240 SNP loci, and carrying out PCR amplification;
(a3) Detecting FAM and HEX fluorescence values of the PCR amplification product of the DNA of the tobacco sample to be tested at the 240 SNP sites by using a KASP fluorescence detector, and calculating to obtain the genotyping result of the tobacco sample to be tested at the 240 SNP sites (namely detecting specific nucleotides at the SNP sites, the same below). Data of a single SNP locus is recorded as XX, YY or XY, wherein X and Y are single nucleotide polymorphisms of the locus, specifically A, T, G or C, and are arranged in the sequence. Missing or invalid data is recorded as- -. If the sum of the XY and- -genotypes of a sample is higher than 10%, it is not used.
The method B comprises the following steps: a method for constructing a DNA fingerprint of a tobacco germplasm resource/variety comprises the following steps (b 1):
(b1) Extracting genome DNA of all tobacco germplasm resources/varieties for constructing the DNA fingerprint, detecting the genotypes of the 240 SNP sites to complete genotyping of the tobacco germplasm resources/varieties to be tested, and combining and arranging the genotypes of each tobacco germplasm resource/variety to be tested on the 240 SNP sites according to the site numbering sequence to form a specific sequence which can be used as the DNA fingerprint of the tobacco germplasm resources/varieties.
Further, the method can further comprise the following step (b 2) on the basis of the step (b 1):
(b2) From the 240 SNP loci mentioned above, based on the principle of 2 SNP loci per chromosome, ts003, ts007, ts013, ts019, ts028, ts029, ts033, ts039, ts041, ts048, ts055, ts060, ts062, ts070, ts074, ts079, ts082, ts089, ts093, ts098, ts106, ts108, ts113, ts120, ts122, ts129, ts132, ts138, ts141, ts148, ts152, ts159, ts164, ts169, ts172, ts179, ts183, ts186, ts192, ts194, ts201, ts207, ts212, ts218, ts222, ts230, ts232 and Ts236 share 48 SNP loci as core SNP characteristic loci for constructing a tobacco germplasm/DNA fingerprint map.
Further, the method can further comprise the following step (b 3) on the basis of the step (b 2):
(b3) Aiming at the 48 core characteristic SNP loci, converting genotype combinations into computer character string labels according to the following steps:
firstly, according to the combination rule of SNP loci on the same chromosome, 48 core characteristic SNP loci are divided into 24 pairs of SNP locus combinations. The genotyping result of each SNP site consists of XX, YY, XY or-wherein X and Y are the mononucleotides of the site, such as A, T, G, C, arranged in this order. Two SNP loci in each pair of SNP locus combinations are arranged in sequence from small to large according to physical positions on a chromosome, and each pair of SNP locus genotype combinations comprise homozygous genotype combinations of AAAA, AATT, AAGG, AACC, TTAA, TTTT, TTGG, TTCC, GGAA, GGTT, GGGGGG, GGCC, CCAA, CCTT, CCGG and CCCC and genotype combinations containing XY or-type.
And then, converting homozygous genotype combinations of each pair of SNP loci according to the character '0' corresponding to AAAA, the character '1' corresponding to AATT, the character '2' corresponding to AAGG, the character '3' corresponding to AACC, the character '4' corresponding to TTAA, the character '5' corresponding to TTTT, the character '6' corresponding to TTGG, the character '7' corresponding to TTCC, the character '8' corresponding to GGAA, the character '9' corresponding to GGTT, the character 'a' corresponding to GGCC, the character 'b' corresponding to CCAA, the character'd' corresponding to CCTT, the character 'e' corresponding to CCGG and the character 'f' corresponding to CCCC. A genotype combination containing XY or- -type is defined as "nonsense combination" and is uniformly converted to the character 'n'. The 24 SNP site combinations are switched according to the chromosome number in turn.
According to the above rules, 48 core characteristic molecule SNP locus genotype combinations in the tobacco germplasm resources/varieties DNA fingerprint map are converted into a character string consisting of 24 characters, and the character string is used as a core code of the tobacco germplasm resources/varieties DNA fingerprint map. The core code of a DNA fingerprint map of a significant tobacco germplasm resource/variety contains no more than 3 characters 'n'.
Further, the method can further comprise the following step (b 4) on the basis of the step (b 3):
(b4) In order to avoid the situation that the genetic relationship is similar to the core code of the tobacco germplasm resource/variety, on the basis of defining the core code of the DNA fingerprint of the tobacco germplasm resource/variety, the 240 SNP locus genotype data are combined, and an additional code and an annotation code section are newly added, wherein the format is as follows: and (c) core code-additional code-annotation code, wherein the core code is a character string formed by 24 characters defined in the step (b 3), the additional code is 1 of the characters 0-9, a-f and n, and the annotation code is a character string formed by the characters 0-9 and with the length of 6.
The additional code is any one of the above-mentioned 240 SNP loci except 48 core characteristic SNP loci which can distinguish the genotype codes of the combination of two SNP loci of the tested tobacco germplasm resources/varieties, and the coding rule is carried out with reference to (b 3).
The annotation code consists of the Arabic numbers in the sequence numbers of the two SNP sites corresponding to the additional codes. If the core codes of the two tobacco germplasm resources/varieties Z1 and Z2 to be tested are the same, but the genotypes of a few SNP loci including Tsxxx and Tsyyy (corresponding to the column of the "sequence number" in Table 2) are different from each other in the 240 SNP loci, the annotation code is 'xxxyyy'.
Further, the method can further comprise the following step (b 5) on the basis of the step (b 4):
(b5) The constructed DNA fingerprint core code and core code-additional code-annotation code characters of the tobacco germplasm resources/varieties are converted into a two-dimensional code or a bar code which is easy to identify by equipment terminals such as a code scanning gun, a mobile phone and the like, so that the tobacco germplasm resources/varieties are intelligently identified and managed.
The method C comprises the following steps: a method for detecting the purity and/or the authenticity of a tobacco variety comprises the following steps:
(c1) Preparing more than 50 seeds of a tobacco variety to be detected and a standard control tobacco variety respectively, accelerating germination and sowing, taking an equivalent tissue of a single seedling when the seedling grows out of 4 true leaves, dividing the seedling into two groups according to the tobacco variety to be detected and the standard control tobacco variety, mixing and preparing samples, and extracting genome DNA;
(c2) And detecting the genotypes of the 240 SNP sites to obtain the genotype data of the tobacco variety to be detected and the standard control tobacco variety on the 240 SNP sites. When the genotype of the standard control tobacco variety sample at a certain SNP locus is XX and YY, the locus is an effective locus, and if the genotype of the locus is XY or-type, the locus is an ineffective locus. If the proportion of the invalid sites of the standard control tobacco variety sample is higher than 10%, the standard control tobacco variety sample is not adopted, and the test is invalid.
(c3) Aiming at the effective locus of the standard control tobacco variety sample in the 240 SNP loci, the genotype difference of the tobacco variety to be detected and the standard control tobacco variety is compared, and the results are two types: the genotypes are the same or different.
(c4) The value of the breed purity (P) is expressed in% and is calculated according to equation (1):
Figure BDA0003265559030000231
in the formula:
p-variety purity;
s1-number of significant loci of said standard control tobacco variety;
s2, in the effective sites of the standard comparison tobacco variety, the number of the sites with different genotypes of the tobacco variety to be detected and the standard comparison tobacco variety is different.
(c5) When the purity of the tobacco variety to be detected is less than or equal to 95%, the tobacco variety to be detected and a standard comparison variety are judged to be different varieties, when the purity of the tobacco variety to be detected is greater than 95% and less than 100%, the tobacco variety to be detected is judged to be an approximate variety or a suspected same variety of the standard comparison variety, and when the purity of the tobacco variety to be detected is 100%, the tobacco variety to be detected and the standard comparison variety are judged to be the same variety.
In the aspects, the tobacco varieties are conventional varieties, and the types can be selected from various types of common tobacco (Nicotina tabacum L.) varieties such as flue-cured tobacco, sun-cured tobacco, burley tobacco, cigar tobacco, aromatic tobacco, drug tobacco and the like.
In a particular embodiment of the invention, said tobacco is selected in particular from the 150 parts of tobacco shown in table 1.
Further, in the method of the fifth aspect, the set of KASP primers of the second aspect or the kit of the third aspect may be used to genotype the 240 SNP sites.
The method is based on more than one million SNP variation site data obtained by the tobacco whole genome re-sequencing work carried out by the Guizhou province tobacco science research institute, the variation site data are further analyzed, 240 SNP markers and matched KASP primers are identified and screened by utilizing a KASP technical platform, and 24 chromosomes of the tobacco are covered. The invention provides a set of credible SNP markers which are convenient to be utilized in later period for the research of tobacco biology, and the set of SNP markers and detection primers can be used for the development of a tobacco SNP detection kit, the construction of a tobacco variety DNA fingerprint, the genotyping of tobacco germplasm resources and genetic groups, the construction of a tobacco germplasm resource/variety DNA fingerprint, the detection of the purity/truth degree of tobacco varieties and the like.
Drawings
FIG. 1 is a data record of the tobacco SNP marker screening process of the present invention.
FIG. 2 is a graph showing the distribution of 240 SNP markers on a tobacco chromosome/linkage group.
FIG. 3 is an illustration of the results of KASP assay.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The examples provided below serve as a guide for further modifications by a person skilled in the art and do not constitute a limitation of the invention in any way.
The experimental procedures in the following examples, unless otherwise indicated, are conventional and are carried out according to the techniques or conditions described in the literature in the field or according to the instructions of the products. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1 screening and development of 240 SNP markers and KASP primers for tobacco
Based on 150 parts of tobacco germplasm resources/varieties (table 1) material whole genome re-sequencing SNP variation data (provided by Guizhou province tobacco science research institute, with the file name of Tobacco. Vcf and the marker number of 1216749), which are completed by Guizhou province tobacco science research institute, 24 chromosomes covering tobacco are screened out by combining mathematical statistics analysis and KASP test verification, 10 of the chromosomes are distributed relatively uniformly, and 240 SNP markers and matched KASP primers of the material can be distinguished. The specific steps for screening and developing SNP markers and KASP primers are as follows:
(1) Data format sorting, converting a re-sequencing variation result file (vcf format) into a ped and map file which can be read by plink software by using vcftools software, and executing a statement:
vcftools--vcf Tobacco.vcf--plink--noweb--out tobacco_plink
plink (v 1.07) software and perl language script format. Pl are used to convert the data format into txt format containing SNP matrix data, wherein each row contains 1 SNP marker, the 1 st column is the SNP marker name, the 2 nd column is the chromosome number, the 3 rd column is the physical position of the marker on the chromosome, and the 4 th column is 1 tobacco variety material from the beginning to the end.
And (3) executing a statement:
plink--file tobacco_plink--transpose--recode--out tobacco_geno
perl format.pl tobacco_geno.tped>tobacco_geno_format.txt
(2) Statistics of SNP Polymorphism Index (PIC), deletion rate (MR), site Heterozygosity Frequency (HF) of each line in tobaco gene format txt were performed using the ptl language script stat _ minor _ morjor.
And (3) executing the statement:
perl stat_minor_mojor.pl tobacco_geno_format.txt>tobacco_geno_stat.txt
(3) And (3) data quality control:
removing SNP sites with HWE greater than 0.05 in Hardy Winberg equilibrium check;
eliminating SNP locus with minimum allelic locus frequency MAF greater than 0.05;
eliminating SNP sites with PIC less than 0.2 or greater than 0.45;
eliminating SNP locus with MR larger than 0.1;
eliminating SNP sites with HF larger than 0.1;
the pLink software (refer to pLink v1.07 help documentation) can be used to knock out SNP sites where HWE and MAF do not meet the requirements.
The number of markers remaining after quality control of the data at this step was 1179154.
(4) And (3) carrying out Haplotype analysis (Haplotype block estimation) on the SNP loci passing through the step (2) by utilizing plink software, naming the SNP loci according to the analysis result, and specifying rules of c + number + hb + number + s + number, such as c1hb14s146, representing chromosome 1/linkage group, 14 th Haplotype block (Haplotype block) and 146 th SNP marker.
(5) KASP Primer design was performed for the SNP site of step (4) with reference to SNP _ Primer _ Pipeline (https:// githu. Com/pinbo/SNP _ Primer _ Pipeline) of Zhang Legione, which retained the number of SNP markers at 217621 after the end of the step.
(6) Combining the steps (3) to (5), and artificially optimizing more than 30 SNP markers from each chromosome, wherein the standard is as follows:
the step (5) is provided with a mark for designing the KASP primer;
a marker preferably located in the CDS (gene coding region);
a maximum of 2 markers per haplotype block;
the markers are distributed relatively evenly across the chromosome.
(7) Randomly selecting 24 parts from 150 parts of tobacco germplasm resources/varieties, and performing test verification on the SNP site genotypes selected in the step (6) and the effectiveness of the synthesized KASP primer, wherein the 150 parts of tobacco germplasm resources/varieties are shown in Table 1 (stored in a tobacco germplasm resources library of the tobacco science research institute of Guizhou province, and the introduction of each material can refer to the Nippon tobacco variety resources (volume one and volume two), the experimental system refers to the technical documents of the British LGC company (specifically, the example 2), if the KASP test genotypes of the randomly selected 24 parts of tobacco germplasm resources/varieties at a certain SNP site are successful, and the genotypes and the re-sequencing results are consistent, the true existence of the SNP site is shown, and the designed KASP primer can be used.
(8) According to the result in (7), the operation of artificial optimization, primer synthesis, experimental verification and the like is repeated again. 1362 SNP markers are finally selected and verified by KASP test, and the result shows that 736 markers are available markers. The selection markers and the number of available markers per chromosome are shown in FIG. 1.
(9) And (3) genotyping 150 tobacco germplasm resources/varieties (table 1) by using the 1362 SNP markers in the step (8), and sorting experimental data, wherein the 240 SNP markers are preferably selected and used for the relevant research of tobacco biology. Preferred criteria are:
each chromosome is divided into 10 sections according to the length on average, and each section is selected from 1 marker as much as possible;
selecting 1 mark in CDS area in each section;
the distance between adjacent markers is not within one haplotype block;
the 240 markers can completely distinguish 150 tobacco germplasm resources/varieties, and the minimum difference number of homozygous SNP between any two varieties is 2.
The positional genotype information of the 240 SNP markers obtained in the present invention is shown in Table 2. Wherein, the physical position of the SNP marker locus is marked based on the tobacco variety Honghuadajinyuan reference genome V1.2 version sequenced by the Chinese tobacco genome plan.
TABLE 1, 150 tobacco germplasm resources/varieties
Figure BDA0003265559030000251
Figure BDA0003265559030000261
Figure BDA0003265559030000271
Figure BDA0003265559030000281
Figure BDA0003265559030000291
Note: except for introduction of the XZ numbered tobacco germplasm resources/varieties in the table, reference is made to monograph Guizhou tobacco variety resources (Vol I) (Vol II), the XZ numbered tobacco germplasm resources/varieties are newly collected tobacco germplasm in the present year, and all the germplasm resources are stored in a Guizhou tobacco science research institute germplasm resource library.
TABLE 2, 240 SNP site-related information
Figure BDA0003265559030000292
Figure BDA0003265559030000301
Figure BDA0003265559030000311
Figure BDA0003265559030000321
Figure BDA0003265559030000331
Figure BDA0003265559030000341
Figure BDA0003265559030000351
To further clarify the characteristics of the above markers, the nucleotide type of each SNP marker site plus the downstream 50bp sequence are shown in Table 3.
TABLE 3 base types of each SNP marker site plus the downstream 50bp sequence
Figure BDA0003265559030000352
Figure BDA0003265559030000361
Figure BDA0003265559030000371
Figure BDA0003265559030000381
Figure BDA0003265559030000391
Figure BDA0003265559030000401
Figure BDA0003265559030000411
Figure BDA0003265559030000421
The distribution of 240 SNP markers on the tobacco chromosome/linkage group is shown in FIG. 2.
The KASP primers developed by the invention for detecting the 240 SNP markers comprise two forward primers and one reverse primer respectively corresponding to each SNP marker. Wherein the forward primer 1 sequence is connected with the FAM fluorescent tag sequence 5' and ' GAAGGTGACCAAGTTCATGCT-3' (SEQ ID No. 721) at the 5' end, and the forward primer 2 sequence is connected with the HEX fluorescent tag sequence 5' and ' GAAGGTCGGAGTCAACGGATT-3' (SEQ ID No. 722). The primers corresponding to each marker of the present invention are shown in Table 4.
TABLE 4 primers for each marker of the invention
Figure BDA0003265559030000422
Figure BDA0003265559030000431
Figure BDA0003265559030000441
Figure BDA0003265559030000451
Figure BDA0003265559030000461
Note: the sequences in the table correspond to tables 2 and 3.
The invention provides a corresponding detection kit for 240 SNP markers of tobacco (see tables 2 and 3). The kit comprises 240 sets of primers (see table 4) synthesized by bio-engineering (shanghai) gmbh, wherein the forward primer 1 and the forward primer 2 have been added with FAM fluorescent sequence and HEX fluorescent sequence, respectively, and the product is labeled as tasip KASP primemix.
The kit also contains KASP 2 × Mastermix, product catalog number KBS-1016-002, from LGC, inc., UK. Wherein, the 2 XKASP Mix consists of a fluorescent probe A, a fluorescent probe B, a quenching probe A and a quenching probe B, high fidelity Taq enzyme, dNTP and Mg 2+ And the like. The nucleotide sequence of the fluorescent probe A is as follows: 5' gaaggtgaccaagttcatgct-3', wherein the 5' end of the 5' gaaggtgaccaagttcatgct-3' is connected with a FAM fluorescent group; the nucleotide sequence of the fluorescent probe B is as follows: 5' gaaggtcggagtcaagtcaacggatt-3 ', wherein a HEX fluorescent group is connected to the 5' terminal; the nucleotide sequence of the quenching probe A is as follows: 5' AGCATGAACTTGGTCACCTTC-3', the 3' end of which is connected with a quenching group BHQ; the nucleotide sequence of the quenching probe B is as follows: 5' AATCCGTTGACTCCGACCTTC-3', the 3' end of which is connected with a quenching group BHQ.
The kit is also provided with MgCl 2 And ddH 2 O, wherein the MgCl 2 Is a product of Beijing Vocko Biotech Co., ltd, the CAS number of the product is 7786-30-3 2 And O is high-pressure sterilization double distilled water.
Example 2 genotyping of tobacco germplasm resources/varieties
The invention provides a method system for genotyping tobacco germplasm resources or genetic group materials (specifically 150 parts of tobacco materials in table 1) by adopting 240 SNP markers (tables 2 and 3) in a step one and combining practical operation by referring to technical documents of LGC company in the UK, and the method comprises the following steps:
(1) Extracting the DNA of the tobacco sample to be detected: putting 200-300 mg of seedlings or leaves of tobacco varieties to be tested into a 1.5ml centrifuge tube, adding liquid nitrogen for full grinding, and adding 700 mu L of CTAB extracting solution preheated at 65 ℃ into each tube; placing the centrifugal tube in a water bath at 65 ℃ for 30 min-1 h, and slightly shaking the centrifugal tube every 10min; cooling for 2min after water bath is finished, adding 0.5mL of chloroform-isoamylol mixed solution, violently shaking and uniformly mixing, and centrifuging for 5-10 min at 12000 rmp; taking the supernatant in a centrifuge tube to a new centrifuge tube, adding 2 times volume of precooled isopropanol, reversing and uniformly mixing, placing at-20 ℃ for precipitation for more than 30min, centrifuging at 12000rmp for 10min, and discarding the supernatant; washing with 75% ethanol for 2 times, centrifuging at 12000rmp for 3min, discarding supernatant, naturally drying precipitate, dissolving in 50 μ L deionized water or TE, detecting concentration, diluting to 100ng/μ L, and immediately using or storing at-20 deg.C.
The step is only a recommendation method, and other methods which can achieve the same effect can also be adopted.
(2) For 240 SNP markers (Table 2 and Table 3), KASP Mastermix and TaSNP KASP Primermix were added in this order to prepare a PCR reaction system, and PCR amplification was carried out. Wherein, the TaSNP KASP Primermix consists of a forward primer 1 with a fluorescent label FAM connected to the 5 'end, a forward primer 2 with a fluorescent label HEX connected to the 5' end and a reverse primer, and the three are connected with ddH 2 O is mixed according to the volume ratio of 12.
The PCR reaction system is shown in Table 5. The PCR reaction conditions are shown in Table 6.
TABLE 5 PCR reaction System
Figure BDA0003265559030000471
TABLE 6 PCR reaction conditions
Figure BDA0003265559030000472
And (3) PCR reaction: 1) The diluted DNA was dispensed into 200. Mu.l 96-well plates and then subjected to RiNIN 8-well discharge gun (model)Pipet-lite XLS 0.5-10ul LTS) pipette 2 μ l into 384-well PCR plates, centrifuge to the bottom for use (manufacturer AXYGEN); 2) Mixing KASPMastermix, taSNP KASP Primermix, ddH 2 O, etc. in the proportions of 2; 3) Subpackaging PCR-mix into each reaction hole according to 4 mul (dry method) and 2 mul (wet method) of each reaction, centrifuging to the bottom, and sealing by using a silica gel cover or a sealing film; 4) The reaction was carried out in a 384-well PCR apparatus (model Veriti) according to the reaction protocol described above TM 384-Well Thermal Cycler) to complete the reaction.
(3) Fluorescence detection and genotyping: and (3) moving out the 384 pore plate after the PCR is finished, removing the silica gel cover or the sealing film after cooling, placing the plate on an SNP (Single nucleotide polymorphism) typing detector (model FLUOstar Omega SNP) to detect FAM and HEX fluorescence values, and analyzing the genotyping result by using KlusterCaller software to obtain the genotype.
As for the genotyping result of a single SNP marker locus, the data is recorded as ' XX ' or ' XY ', ' YY ', ' XX ' and ' YY ' represent homozygosity ', and ' XY ' represents heterozygosity, wherein X and Y are single nucleotides of the locus, such as A, T, G and C. If the condition that the single nucleotides A, T, G and C cannot be determined appears in a certain SNP marker, the SNP marker is invalid data, and the invalid data is recorded as "-".
FIG. 3 is an exemplary diagram of KASP test results. Black (NTC); red (YY); blue (XX); green (XY). Pink (-).
The proportion of invalid data of one mark is higher than 10%, the mark is invalid mark, the proportion of invalid data of one sample is higher than 10%, and the sample is invalid sample and is not adopted.
(4) 150 tobacco germplasm resources/variety (Table 1) materials were genotyped at the 240 marker loci, with the results shown in Table 7.
TABLE 7, 150 tobacco germplasm resources/variety materials genotyping results
Figure BDA0003265559030000481
Figure BDA0003265559030000491
Figure BDA0003265559030000501
Figure BDA0003265559030000511
Figure BDA0003265559030000521
Figure BDA0003265559030000531
Figure BDA0003265559030000541
Figure BDA0003265559030000551
Figure BDA0003265559030000561
Figure BDA0003265559030000571
Figure BDA0003265559030000581
Figure BDA0003265559030000591
Figure BDA0003265559030000601
Note: the results of typing 240 SNP markers in the table are represented as a string of 480 in length consisting of the characters ` A ` T ` G ` C `, and the string is divided into 240 parts in length of 2, each part representing the genotype of one SNP site, and the order of the sites is arranged in the order of the site numbers in Table 2 from small to large.
Example 3, 150 parts of tobacco germplasm resources/variety DNA fingerprint core code construction
The invention constructs a fingerprint spectrum for 150 tobacco germplasm resources (table 1) stored by Nicotiana tabacum scientific research institute in Guizhou province by using the proposed method for the fingerprint spectrum of the tobacco germplasm resources/varieties, and draws a core code of the fingerprint spectrum, and the method comprises the following steps:
(1) According to the steps of example 2, 150 tobacco germplasm resources/varieties (table 1) are genotyped at the 240 SNP marker sites (table 2 and table 3), and the result is shown in table 7, wherein the result of genotyping the 240 SNP sites corresponding to each material in table 7 is one of the fingerprint representation modes.
(2) And extracting 150 parts of materials to obtain genotypes on 48 core characteristic SNP sites of Ts003, ts007, ts013, ts019, ts028, ts029, ts033, ts039, ts041, ts048, ts055, ts060, ts062, ts070, ts074, ts079, ts082, ts089, ts093, ts098, ts106, ts108, ts113, ts120, ts122, ts129, ts132, ts138, ts141, ts148, ts152, ts159, ts164, ts169, ts172, ts179, ts183, ts186, ts192, ts194, ts201, ts207, ts212, ts218, ts222, ts230, ts232 and Ts236.
(3) The 48 core characteristic SNP marker loci are divided into 24 pairs of SNP locus combinations according to the combination rule of SNP loci on the same chromosome for the genotypes of the 48 loci of a single tobacco germplasm resource/variety material in the step (2). The genotyping result of each SNP site consists of XX, YY, XY or-wherein X and Y are the mononucleotides of the site, such as A, T, G, C, arranged in this order. Two SNP loci in each SNP locus combination are arranged in sequence from small to large according to physical positions on a chromosome, and the genotype combinations of each SNP locus comprise homozygous genotype combinations such as AAAA, AATT, AAGG, AACC, TTAA, TTTT, TTGG, TTCC, GGAA, GGTT, GGGGGG, GGCC, CCAA, CCTT, CCGG, CCCC and the like and genotype combinations containing XY or-type.
And then, converting homozygous genotype combinations of each pair of SNP loci according to the character '0' corresponding to AAAA, the character '1' corresponding to AATT, the character '2' corresponding to AAGG, the character '3' corresponding to AACC, the character '4' corresponding to TTAA, the character '5' corresponding to TTTT, the character '6' corresponding to TTGG, the character '7' corresponding to TTCC, the character '8' corresponding to GGAA, the character '9' corresponding to GGTT, the character 'a' corresponding to GGCC, the character 'b' corresponding to CCAA, the character'd' corresponding to CCTT, the character 'e' corresponding to CCGG and the character 'f' corresponding to CCCC. A genotype combination containing XY or- -type is defined as a "nonsense combination", and is uniformly converted to the character 'n'. The 24 pairs of SNP site combinations are sequentially switched according to the chromosome number.
After the completion, 48 core characteristic SNP locus genotype combinations in the tobacco germplasm resources/variety DNA fingerprint are converted into a character string consisting of 24 characters, and the character string is used as a core code of the tobacco germplasm resources/variety DNA fingerprint.
(4) And sequentially obtaining the core codes of the DNA fingerprint of the 150 tobacco germplasm resources/varieties, wherein the core codes can be converted into two-dimensional codes (shown in table 8) to serve as the characteristics of the corresponding tobacco germplasm resources/varieties and be used for the basic standards of the exchange and selling process of the germplasm resources/varieties.
Table 8, 150 core codes of tobacco germplasm resources/variety DNA fingerprint spectrums and two-dimensional codes converted from core codes
Figure BDA0003265559030000611
Figure BDA0003265559030000621
Figure BDA0003265559030000631
Figure BDA0003265559030000641
Figure BDA0003265559030000651
Figure BDA0003265559030000661
Figure BDA0003265559030000671
Figure BDA0003265559030000681
Figure BDA0003265559030000691
Figure BDA0003265559030000701
Figure BDA0003265559030000711
Figure BDA0003265559030000721
Figure BDA0003265559030000731
Figure BDA0003265559030000741
Figure BDA0003265559030000751
Figure BDA0003265559030000761
The method is based on 150 parts of tobacco whole genome re-sequencing data, utilizes a data analysis method of early exploration to mine and obtain more than one million SNP sites, further analyzes the sites, utilizes a KASP technical platform to identify and screen 240 SNP markers and matched KASP primers, and covers 24 chromosomes/linkage groups of tobacco. The invention provides a set of credible SNP markers which are convenient to be utilized in later period for the research of tobacco biology, and the set of SNP markers and detection primers can be used for developing a tobacco SNP detection kit, constructing a tobacco variety DNA fingerprint, genotyping tobacco germplasm resources and genetic groups, detecting the purity/authenticity of the tobacco variety and the like.
The present invention has been described in detail above. It will be apparent to those skilled in the art that the invention can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation. While the invention has been described with reference to specific examples, it will be appreciated that the invention may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains. The use of some of the essential features is made possible within the scope of the claims attached below.
<110> Guizhou province tobacco science research institute
<120> tobacco SNP marker developed based on whole genome re-sequencing and KASP technology and application thereof
<130> GNCLN211246
<160> 722
<170> PatentIn version 3.5
<210> 1
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 1
cctgtgacaa gtaaagcttt ctttt 25
<210> 2
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 2
cagatcatcg tgtaacctcc tttt 24
<210> 3
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 3
cgttattatg tcaagaagat gcaga 25
<210> 4
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 4
atgatgaatc gccggcgga 19
<210> 5
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 5
tgatgcagat ttagctcaga atca 24
<210> 6
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 6
ggagcttgat gaggtaatcc at 22
<210> 7
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 7
tttcccagat acccgcccat 20
<210> 8
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 8
tcgcgcaata ttgtacttct ga 22
<210> 9
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 9
cgtcacgata attcacagag gga 23
<210> 10
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 10
tcccacgtgt caagttgaat t 21
<210> 11
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 11
aatccaccca cgatgcctat 20
<210> 12
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 12
cttggatggt ttgaggccaa 20
<210> 13
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 13
tttgcttctg ctggaatcga 20
<210> 14
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 14
ctggaacgcc ggtgatggt 19
<210> 15
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 15
tgtgaataat ggttgcaaag tgtt 24
<210> 16
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 16
aggatcttat ggtcgaaagt caa 23
<210> 17
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 17
tggctagtct ctagagcaca t 21
<210> 18
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 18
acgagaatag gtttggagtg ttt 23
<210> 19
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 19
gtcgatacct tcttggccca 20
<210> 20
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 20
cagcttcgga taaaagttgg act 23
<210> 21
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 21
accttatctg ccaaaggttc aca 23
<210> 22
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 22
tgggcgaggc tcatggat 18
<210> 23
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 23
cgaaccccat cactttcttg aat 23
<210> 24
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 24
tcctgcttct ttgtacttgt acca 24
<210> 25
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 25
tgtgaatact aggttgcggg a 21
<210> 26
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 26
tccgctagaa tcggtccgaa 20
<210> 27
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 27
gccatcctca aaatcaagtc catt 24
<210> 28
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 28
ggtataccag gaacaacgcc a 21
<210> 29
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 29
tcctacttca cttccatttt actta 25
<210> 30
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 30
cagcatctgt tccagcttct 20
<210> 31
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 31
agctcctgaa tctgctcaac t 21
<210> 32
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 32
gtgaaaagcg cttccaggaa 20
<210> 33
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 33
tcagagtttg gacttgatgt cc 22
<210> 34
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 34
ggcctcgcta ctcttcga 18
<210> 35
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 35
gctctgtctt agtcacttgc ttc 23
<210> 36
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 36
ctgggcgttt atcactcact 20
<210> 37
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 37
gttcgacccc tacctggt 18
<210> 38
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 38
aacctgcaat tgctgatgca 20
<210> 39
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 39
tcaataacca catcgtgaaa ccg 23
<210> 40
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 40
agatatgacg gctgagtaca ac 22
<210> 41
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 41
agaaggttct gagctgtcta cttc 24
<210> 42
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 42
ggggctaccg tcattgatta a 21
<210> 43
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 43
ccatttcttc caaagctaaa gttgt 25
<210> 44
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 44
aggtccacga agtattggca 20
<210> 45
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 45
atgttgtgac ttctcttggg att 23
<210> 46
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 46
acggccgtca tggttggt 18
<210> 47
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 47
tgtcactggg tgaaaatcca tca 23
<210> 48
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 48
tcaaggctta gcttgtcata gttta 25
<210> 49
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 49
tcattctctc tctctctttc gagaa 25
<210> 50
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 50
tgggtacacg ttgctttcgt 20
<210> 51
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 51
cagaatatgg gaatttcggg tctta 25
<210> 52
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 52
acaggagcag ttgaataagg cta 23
<210> 53
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 53
ggatacttta tgcttgcctt atgat 25
<210> 54
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 54
accactcgtc tttgactcaa tca 23
<210> 55
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 55
tgttttcttt acagcggata atgtt 25
<210> 56
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 56
agatagaact tgttgatgat ttgca 25
<210> 57
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 57
tggaattcca tgagtttgga tgtt 24
<210> 58
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 58
gacttcaaca gtcgcgactt 20
<210> 59
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 59
gaggaaggat catcgccgaa ta 22
<210> 60
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 60
ggagggagat tctgcagact 20
<210> 61
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 61
gaagagtcaa tcatcgatga atcta 25
<210> 62
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 62
ccgttgcaac tttaattcca aaatt 25
<210> 63
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 63
gctgcaggtt gagttaatcc atat 24
<210> 64
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 64
acccctgaaa atttaaacga ttcat 25
<210> 65
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 65
cagctacaac cggtggtact a 21
<210> 66
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 66
acagagcctt cttgaaggag at 22
<210> 67
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 67
ctaccttcgc ggcggagat 19
<210> 68
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 68
acattttgca gggagtcact tc 22
<210> 69
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 69
ctgggcggag gtagttctt 19
<210> 70
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 70
accctaatcc ttcacttacc aga 23
<210> 71
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 71
ttgcggcaac aatctgagat 20
<210> 72
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 72
ccattaagga caagctcgaa gata 24
<210> 73
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 73
ggttttggtt tacctaagtg gagt 24
<210> 74
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 74
cgctaaggat tacaatattg aagga 25
<210> 75
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 75
agagcctcac tttcctggat 20
<210> 76
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 76
gatagcccat gtgcgagagt 20
<210> 77
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 77
gctgcctctg cgataaatct 20
<210> 78
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 78
acgacgtgtc catggcct 18
<210> 79
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 79
agaaaataaa gcgcaaaggc aaag 24
<210> 80
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 80
tctgctccct catatacccc t 21
<210> 81
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 81
tcctaccctc aaagaaaagg tca 23
<210> 82
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 82
agatcctgtc aaaatctgag gaaga 25
<210> 83
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 83
gtgctggagg gcattgca 18
<210> 84
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 84
tcccaaatgg tccctcagga 20
<210> 85
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 85
ccaagaacag gcaaaacca 19
<210> 86
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 86
ttgatatcag aggattctcg agaga 25
<210> 87
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 87
gaacatggct ccttgcatgt 20
<210> 88
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 88
gctacagcac ctgcacatgt 20
<210> 89
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 89
tgacatgtga ggttgactct cta 23
<210> 90
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 90
gagtctcact ctattctttc tcgaa 25
<210> 91
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 91
ccagtgccca gttatacaac act 23
<210> 92
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 92
tctcttccaa gtgcccatga 20
<210> 93
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 93
tcgtgttaac cttcctcgga 20
<210> 94
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 94
agggatgtct tcatagaaag gctt 24
<210> 95
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 95
tgattcaaga gcttccttca gt 22
<210> 96
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 96
agaatcgggt ttttgaaggc c 21
<210> 97
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 97
acttaggatg tttagccatt ttga 24
<210> 98
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 98
agaccttcaa atgttcgcca tta 23
<210> 99
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 99
gtgccacgtg tgccaatta 19
<210> 100
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 100
agcaaaattt acctgtgggg a 21
<210> 101
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 101
ctgtacttgg tgcttggcta ta 22
<210> 102
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 102
ctcgtctctg tcgatcgtca t 21
<210> 103
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 103
ccctttttcc ctcctcgca 19
<210> 104
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 104
tgctccaagc tctccttcaa 20
<210> 105
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 105
gaagagtgag tttgaccttg aaatt 25
<210> 106
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 106
tctccaaatc cattctcagt cctt 24
<210> 107
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 107
actcttcgcg aacgcaaagt 20
<210> 108
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 108
tcgtcgccta aaatcgtgca 20
<210> 109
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 109
ggagcaatta gtggctctgt 20
<210> 110
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 110
catttccgat aatttggtca attcc 25
<210> 111
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 111
catgcctata cgttcgcaag a 21
<210> 112
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 112
tcgccaatgg gaattttaca ttg 23
<210> 113
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 113
acactcggcc catttctcaa 20
<210> 114
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 114
tgtgtgtctt gtttctcctc ta 22
<210> 115
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 115
tcaacgaaag agacacctgt ta 22
<210> 116
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 116
agcatcctca ttatccacac caat 24
<210> 117
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 117
gaacaggacg gagggttcgt 20
<210> 118
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 118
gcttccccgt ttctaggcta 20
<210> 119
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 119
gcttcaagaa gtttacgact attga 25
<210> 120
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 120
aatggatact cgaccccagt 20
<210> 121
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 121
tcgaaaaatt acagaacgcg att 23
<210> 122
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 122
agggcaagcg ttcattgaat g 21
<210> 123
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 123
ccaaacatct ttcacggcca 20
<210> 124
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 124
aaggatgagt ggagctgtgc 20
<210> 125
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 125
tgtgggaatt cagaacagga t 21
<210> 126
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 126
cattgaaatt gcgaaacttt ttctt 25
<210> 127
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 127
gtcttctgac tcggacattg aaaa 24
<210> 128
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 128
actctctctc acctgttttg caa 23
<210> 129
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 129
acgagttacc atgcttgcaa ac 22
<210> 130
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 130
cgtacacctg aattgacagt ct 22
<210> 131
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 131
taggtttggc caagttggaa 20
<210> 132
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 132
cactggtggc aattggattg t 21
<210> 133
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 133
ccaatgtacc taggatcaaa catat 25
<210> 134
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 134
ggcaggagga gcttctgat 19
<210> 135
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 135
atcctacctt gctctgaaga ta 22
<210> 136
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 136
accgtcatat tagaccacgc t 21
<210> 137
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 137
cccgactcat tgttgtatag atcat 25
<210> 138
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 138
tctcttcccg gcaataatgg t 21
<210> 139
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 139
cttgctgaaa cgccagtatc t 21
<210> 140
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 140
acaggatgga gtcacaagca t 21
<210> 141
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 141
accacaaatg ttgttcaaga attgt 25
<210> 142
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 142
tcaatctctg gaacttcaaa aacat 25
<210> 143
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 143
ggagaacgac tctgaggaag aa 22
<210> 144
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 144
gcacaacatt ttctacaata atggt 25
<210> 145
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 145
cccgtttgaa atagcactat tcctt 25
<210> 146
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 146
actttggaag aaatgcgagt cg 22
<210> 147
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 147
atcacgaaat gcaatgtcca tt 22
<210> 148
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 148
agcttgaacc atccattgtt gca 23
<210> 149
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 149
tggtgcagat gtagctgaaa gt 22
<210> 150
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 150
aatggaactc aatttgcagc a 21
<210> 151
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 151
gcatgaacat gtctatgatg ctgt 24
<210> 152
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 152
cgttactgag gcaaatttca ttgaa 25
<210> 153
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 153
aggaaaccgg agtcaacttt 20
<210> 154
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 154
cgaatgagat gggttagtgg tttt 24
<210> 155
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 155
gcccgaatcg atcagaaaca tt 22
<210> 156
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 156
aacgcgttcc cttcctcaca 20
<210> 157
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 157
aaggacaaga gccgcagagt 20
<210> 158
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 158
ggattggatt aggtaacatc atgag 25
<210> 159
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 159
cttccacaga agcggaatat tt 22
<210> 160
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 160
ccaaaatact gctgcaccgt 20
<210> 161
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 161
actggaacta gagcctcatc a 21
<210> 162
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 162
acgttcacca aagaggatca 20
<210> 163
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 163
agtcgctctt cggtctaatt ctt 23
<210> 164
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 164
aaaaagggcc atggactcca 20
<210> 165
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 165
gcaacctgcg ctgaacat 18
<210> 166
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 166
ccaatgaaga atccatccac tagt 24
<210> 167
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 167
gttacaactt gccaaccagg at 22
<210> 168
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 168
agcgagtgag agcctttcat ata 23
<210> 169
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 169
tctgctttag cctagatttc acca 24
<210> 170
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 170
aagtttttga agaaggcaat aatga 25
<210> 171
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 171
aagctttaat cacagcagat aaagt 25
<210> 172
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 172
cgtgccgttc gaacctcta 19
<210> 173
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 173
cacacatctt cttcacaagc tca 23
<210> 174
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 174
ccaagatgtt gagaatatgg ctgat 25
<210> 175
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 175
tggaaccaac atcaagtagc ca 22
<210> 176
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 176
agcatctcca agtattgaag cta 23
<210> 177
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 177
ctgaccccct tgctcaca 18
<210> 178
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 178
tgattgtgga taaagtgctc tgtaa 25
<210> 179
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 179
tctacgtttc tgcggactga 20
<210> 180
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 180
cggattcagc agattctttt cttt 24
<210> 181
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 181
actaggagat ctggcacgta at 22
<210> 182
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 182
cgtctttatg aggtcaatcg ga 22
<210> 183
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 183
ttgaattacc agacactaaa tgct 24
<210> 184
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 184
gtcatgacag ttgacgacct cctg 24
<210> 185
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 185
acaacaacat acccagtgaa attt 24
<210> 186
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 186
tcaattcagc tccattgaat tcgt 24
<210> 187
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 187
actctcccaa attaagcttt cctt 24
<210> 188
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 188
cctgaatcgt ttaactcgtt ctca 24
<210> 189
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 189
ctagatctca ggaaactcaa ggtaa 25
<210> 190
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 190
ggaaacgttc atggcaatta ttttt 25
<210> 191
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 191
cacccctttg tcatcttcgt tt 22
<210> 192
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 192
cacttccctc acaactgatg t 21
<210> 193
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 193
tgaattcact ccttagacct caata 25
<210> 194
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 194
tctagtgcgt tcttacttct cat 23
<210> 195
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 195
tgaagtagat caaattggcc gt 22
<210> 196
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 196
ctatctccga tataggggta aagag 25
<210> 197
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 197
aaccatgcac tgggtagttc tt 22
<210> 198
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 198
gacggaagaa gaacagttag ga 22
<210> 199
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 199
gagcccctac tccgtgatct 20
<210> 200
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 200
tctatggctt tgaatgataa gcagt 25
<210> 201
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 201
gcagcaacag cagatggagt 20
<210> 202
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 202
cgtggactat ggaggtcgaa 20
<210> 203
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 203
tgagcaatac atttctgttg tagt 24
<210> 204
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 204
gctacctttc ttagctggga tcat 24
<210> 205
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 205
acgcaggtgc agaccaaa 18
<210> 206
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 206
cgtccttgtc tgtttaaagc tca 23
<210> 207
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 207
tggccaagag tccagacatt 20
<210> 208
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 208
cctcaccact tgcaacataa tgt 23
<210> 209
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 209
tgaaataaat gaccctttct cctca 25
<210> 210
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 210
gcattgctgg gtcatcattc t 21
<210> 211
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 211
ttctgtcaat atctttcttc agcta 25
<210> 212
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 212
accttcttct tcctccaatc cat 23
<210> 213
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 213
cgcatttggg atcaggttct 20
<210> 214
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 214
aaaaggtcat cacaaaccaa aaat 24
<210> 215
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 215
tccaatgaat acaggatggt ca 22
<210> 216
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 216
ccaagtatct aattagttca gccta 25
<210> 217
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 217
ggactcaaga gaatcatcat tgata 25
<210> 218
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 218
tgacgctgtt aatcaattgc tcc 23
<210> 219
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 219
agtaagtcca gttccttcag tca 23
<210> 220
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 220
aattttggtg ccactccatt a 21
<210> 221
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 221
tgtcctaagt gctcatcatc ct 22
<210> 222
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 222
ttctaacaat ctcatcaacg acatt 25
<210> 223
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 223
agctgtggtc atattatgca atgtt 25
<210> 224
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 224
gaggtttctc taatgcacaa gcact 25
<210> 225
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 225
tgcccctaac cagtaacttg t 21
<210> 226
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 226
ggctggcttt ttatatccaa ggt 23
<210> 227
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 227
cctagctcgg tttccagca 19
<210> 228
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 228
cgaggattag tggcttttga aca 23
<210> 229
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 229
taaggcttca tcctcctggt 20
<210> 230
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 230
gaatctcggg ctcaaagtca tta 23
<210> 231
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 231
ccagagtcaa cgatacctac gt 22
<210> 232
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 232
ttcctcttca tcccgttgat t 21
<210> 233
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 233
caaactgata tacactcacc tcaaa 25
<210> 234
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 234
cgcgactgtc ggctgact 18
<210> 235
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 235
tgcactacta gatggtggca taa 23
<210> 236
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 236
agttcacctt ctctacagtt tcct 24
<210> 237
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 237
cgtccgcaga ccttgaagtt ata 23
<210> 238
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 238
cagccacagc catgctca 18
<210> 239
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 239
agtagggtac tcattccata attca 25
<210> 240
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 240
cagagtccag atggctatac tgtg 24
<210> 241
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 241
cctgtgacaa gtaaagcttt ctttc 25
<210> 242
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 242
cagatcatcg tgtaacctcc tttc 24
<210> 243
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 243
cgttattatg tcaagaagat gcagg 25
<210> 244
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 244
atgatgaatc gccggcggc 19
<210> 245
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 245
tgatgcagat ttagctcaga atcg 24
<210> 246
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 246
ggagcttgat gaggtaatcc ac 22
<210> 247
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 247
tttcccagat acccgcccac 20
<210> 248
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 248
tcgcgcaata ttgtacttct gg 22
<210> 249
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 249
cgtcacgata attcacagag ggg 23
<210> 250
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 250
tcccacgtgt caagttgaat a 21
<210> 251
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 251
aatccaccca cgatgcctac 20
<210> 252
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 252
cttggatggt ttgaggccat 20
<210> 253
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 253
tttgcttctg ctggaatcgg 20
<210> 254
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 254
ctggaacgcc ggtgatggc 19
<210> 255
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 255
tgtgaataat ggttgcaaag tgtc 24
<210> 256
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 256
aggatcttat ggtcgaaagt cag 23
<210> 257
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 257
tggctagtct ctagagcaca c 21
<210> 258
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 258
acgagaatag gtttggagtg ttc 23
<210> 259
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 259
gtcgatacct tcttggcccg 20
<210> 260
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 260
cagcttcgga taaaagttgg acc 23
<210> 261
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 261
accttatctg ccaaaggttc acg 23
<210> 262
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 262
tgggcgaggc tcatggac 18
<210> 263
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 263
cgaaccccat cactttcttg aac 23
<210> 264
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 264
tcctgcttct ttgtacttgt accg 24
<210> 265
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 265
tgtgaatact aggttgcggg g 21
<210> 266
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 266
tccgctagaa tcggtccgag 20
<210> 267
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 267
gccatcctca aaatcaagtc catg 24
<210> 268
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 268
ggtataccag gaacaacgcc g 21
<210> 269
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 269
tcctacttca cttccatttt acttg 25
<210> 270
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 270
cagcatctgt tccagcttcc 20
<210> 271
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 271
agctcctgaa tctgctcaac c 21
<210> 272
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 272
gtgaaaagcg cttccaggag 20
<210> 273
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 273
tcagagtttg gacttgatgt cg 22
<210> 274
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 274
ggcctcgcta ctcttcgg 18
<210> 275
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 275
gctctgtctt agtcacttgc ttg 23
<210> 276
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 276
ctgggcgttt atcactcacc 20
<210> 277
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 277
gttcgacccc tacctggc 18
<210> 278
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 278
aacctgcaat tgctgatgcg 20
<210> 279
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 279
tcaataacca catcgtgaaa ccc 23
<210> 280
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 280
agatatgacg gctgagtaca ag 22
<210> 281
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 281
accaacaacc tccttcctca tg 22
<210> 282
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 282
atccactctc caacttcaac tg 22
<210> 283
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 283
gctagctctt ggggttgaga tg 22
<210> 284
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 284
agccggataa gtttacgtat gg 22
<210> 285
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 285
acagtcagcc tggagatttt tac 23
<210> 286
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 286
gcattgcgag ttttaccaat gtttc 25
<210> 287
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 287
cctttctggg gtggcgat 18
<210> 288
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 288
gtgtcctttc cagagtaaca tgg 23
<210> 289
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 289
tcattctctc tctctctttc gagat 25
<210> 290
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 290
tgggtacacg ttgctttcga 20
<210> 291
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 291
cagaatatgg gaatttcggg tcttg 25
<210> 292
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 292
acaggagcag ttgaataagg ctg 23
<210> 293
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 293
ggatacttta tgcttgcctt atgac 25
<210> 294
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 294
accactcgtc tttgactcaa tcg 23
<210> 295
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 295
tgttttcttt acagcggata atgtc 25
<210> 296
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 296
agatagaact tgttgatgat ttgcg 25
<210> 297
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 297
tggaattcca tgagtttgga tgtg 24
<210> 298
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 298
gacttcaaca gtcgcgacta 20
<210> 299
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 299
gaggaaggat catcgccgaa tg 22
<210> 300
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 300
ggagggagat tctgcagacc 20
<210> 301
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 301
gaagagtcaa tcatcgatga atctg 25
<210> 302
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 302
ccgttgcaac tttaattcca aaatg 25
<210> 303
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 303
gctgcaggtt gagttaatcc atac 24
<210> 304
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 304
acccctgaaa atttaaacga ttcag 25
<210> 305
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 305
cagctacaac cggtggtact g 21
<210> 306
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 306
acagagcctt cttgaaggag aa 22
<210> 307
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 307
ctaccttcgc ggcggagac 19
<210> 308
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 308
acattttgca gggagtcact tg 22
<210> 309
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 309
ctgggcggag gtagttctg 19
<210> 310
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 310
accctaatcc ttcacttacc agt 23
<210> 311
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 311
ttgcggcaac aatctgagac 20
<210> 312
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 312
ccattaagga caagctcgaa gatg 24
<210> 313
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 313
ggttttggtt tacctaagtg gagc 24
<210> 314
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 314
cgctaaggat tacaatattg aaggg 25
<210> 315
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 315
agagcctcac tttcctggag 20
<210> 316
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 316
gatagcccat gtgcgagagg 20
<210> 317
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 317
gctgcctctg cgataaatcc 20
<210> 318
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 318
acgacgtgtc catggccc 18
<210> 319
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 319
agaaaataaa gcgcaaaggc aaac 24
<210> 320
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 320
tctgctccct catatacccc c 21
<210> 321
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 321
tcctaccctc aaagaaaagg tct 23
<210> 322
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 322
agatcctgtc aaaatctgag gaagg 25
<210> 323
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 323
gtgctggagg gcattgcg 18
<210> 324
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 324
tcccaaatgg tccctcaggc 20
<210> 325
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 325
ccaagaacag gcaaaaccg 19
<210> 326
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 326
ttgatatcag aggattctcg agagg 25
<210> 327
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 327
gaacatggct ccttgcatgc 20
<210> 328
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 328
gctacagcac ctgcacatgc 20
<210> 329
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 329
tgacatgtga ggttgactct ctg 23
<210> 330
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 330
gagtctcact ctattctttc tcgag 25
<210> 331
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 331
ccagtgccca gttatacaac acg 23
<210> 332
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 332
tctcttccaa gtgcccatgg 20
<210> 333
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 333
tcgtgttaac cttcctcggg 20
<210> 334
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 334
agggatgtct tcatagaaag gcta 24
<210> 335
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 335
tgattcaaga gcttccttca gc 22
<210> 336
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 336
agaatcgggt ttttgaaggc g 21
<210> 337
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 337
acttaggatg tttagccatt ttgg 24
<210> 338
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 338
agaccttcaa atgttcgcca ttg 23
<210> 339
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 339
gtgccacgtg tgccaattg 19
<210> 340
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 340
agcaaaattt acctgtgggg g 21
<210> 341
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 341
ctgtacttgg tgcttggcta tg 22
<210> 342
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 342
ctcgtctctg tcgatcgtca c 21
<210> 343
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 343
ccctttttcc ctcctcgcg 19
<210> 344
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 344
tgctccaagc tctccttcag 20
<210> 345
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 345
gaagagtgag tttgaccttg aaatg 25
<210> 346
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 346
tctccaaatc cattctcagt cctg 24
<210> 347
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 347
actcttcgcg aacgcaaagg 20
<210> 348
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 348
tcgtcgccta aaatcgtgct 20
<210> 349
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 349
aggtggggct tgcattacaa 20
<210> 350
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 350
catttccgat aatttggtca attct 25
<210> 351
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 351
catgcctata cgttcgcaag g 21
<210> 352
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 352
tcgccaatgg gaattttaca ttc 23
<210> 353
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 353
acactcggcc catttctcac 20
<210> 354
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 354
tgtgtgtctt gtttctcctc tc 22
<210> 355
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 355
tcaacgaaag agacacctgt tg 22
<210> 356
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 356
agcatcctca ttatccacac caac 24
<210> 357
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 357
gaacaggacg gagggttcgc 20
<210> 358
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 358
gcttccccgt ttctaggctg 20
<210> 359
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 359
gcttcaagaa gtttacgact attgc 25
<210> 360
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 360
aatggatact cgaccccagc 20
<210> 361
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 361
tcgaaaaatt acagaacgcg atc 23
<210> 362
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 362
agggcaagcg ttcattgaat c 21
<210> 363
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 363
ccaaacatct ttcacggcct 20
<210> 364
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 364
aaggatgagt ggagctgtgg 20
<210> 365
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 365
tgtgggaatt cagaacagga c 21
<210> 366
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 366
cattgaaatt gcgaaacttt ttctg 25
<210> 367
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 367
gtcttctgac tcggacattg aaag 24
<210> 368
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 368
actctctctc acctgttttg cag 23
<210> 369
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 369
acgagttacc atgcttgcaa ag 22
<210> 370
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 370
cgtacacctg aattgacagt cc 22
<210> 371
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 371
taggtttggc caagttggag 20
<210> 372
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 372
cactggtggc aattggattg c 21
<210> 373
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 373
ccaatgtacc taggatcaaa catac 25
<210> 374
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 374
ggcaggagga gcttctgac 19
<210> 375
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 375
atcctacctt gctctgaaga tg 22
<210> 376
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 376
accgtcatat tagaccacgc c 21
<210> 377
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 377
cccgactcat tgttgtatag atcag 25
<210> 378
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 378
tctcttcccg gcaataatgg c 21
<210> 379
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 379
cttgctgaaa cgccagtatc a 21
<210> 380
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 380
acaggatgga gtcacaagca c 21
<210> 381
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 381
accacaaatg ttgttcaaga attgc 25
<210> 382
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 382
tcaatctctg gaacttcaaa aacac 25
<210> 383
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 383
ggagaacgac tctgaggaag ag 22
<210> 384
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 384
gcacaacatt ttctacaata atggg 25
<210> 385
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 385
cccgtttgaa atagcactat tcctg 25
<210> 386
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 386
actttggaag aaatgcgagt cc 22
<210> 387
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 387
atcacgaaat gcaatgtcca tc 22
<210> 388
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 388
agcttgaacc atccattgtt gcc 23
<210> 389
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 389
tggtgcagat gtagctgaaa gc 22
<210> 390
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 390
aatggaactc aatttgcagc c 21
<210> 391
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 391
gcatgaacat gtctatgatg ctgc 24
<210> 392
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 392
cgttactgag gcaaatttca ttgag 25
<210> 393
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 393
aggaaaccgg agtcaacttc 20
<210> 394
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 394
cgaatgagat gggttagtgg tttg 24
<210> 395
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 395
gcccgaatcg atcagaaaca tc 22
<210> 396
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 396
aacgcgttcc cttcctcacg 20
<210> 397
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 397
aaggacaaga gccgcagagc 20
<210> 398
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 398
ggattggatt aggtaacatc atgac 25
<210> 399
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 399
cttccacaga agcggaatat tc 22
<210> 400
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 400
ccaaaatact gctgcaccgg 20
<210> 401
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 401
actggaacta gagcctcatc g 21
<210> 402
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 402
acgttcacca aagaggatcg 20
<210> 403
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 403
agtcgctctt cggtctaatt ctc 23
<210> 404
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 404
aaaaagggcc atggactcct 20
<210> 405
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 405
gcaacctgcg ctgaacac 18
<210> 406
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 406
ccaatgaaga atccatccac tagc 24
<210> 407
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 407
gttacaactt gccaaccagg ac 22
<210> 408
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 408
agcgagtgag agcctttcat att 23
<210> 409
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 409
tctgctttag cctagatttc acct 24
<210> 410
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 410
aagtttttga agaaggcaat aatgg 25
<210> 411
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 411
aagctttaat cacagcagat aaagg 25
<210> 412
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 412
cgtgccgttc gaacctctg 19
<210> 413
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 413
cacacatctt cttcacaagc tcg 23
<210> 414
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 414
ccaagatgtt gagaatatgg ctgag 25
<210> 415
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 415
tggaaccaac atcaagtagc ct 22
<210> 416
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 416
agcatctcca agtattgaag ctc 23
<210> 417
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 417
ctgaccccct tgctcacc 18
<210> 418
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 418
tgattgtgga taaagtgctc tgtag 25
<210> 419
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 419
tctacgtttc tgcggactgt 20
<210> 420
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 420
cggattcagc agattctttt cttc 24
<210> 421
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 421
actaggagat ctggcacgta aa 22
<210> 422
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 422
cgtctttatg aggtcaatcg gg 22
<210> 423
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 423
ttgaattacc agacactaaa tgcc 24
<210> 424
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 424
gtcatgacag ttgacgacct cctc 24
<210> 425
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 425
acaacaacat acccagtgaa attc 24
<210> 426
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 426
tcaattcagc tccattgaat tcgc 24
<210> 427
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 427
actctcccaa attaagcttt cctc 24
<210> 428
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 428
cctgaatcgt ttaactcgtt ctcg 24
<210> 429
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 429
ctagatctca ggaaactcaa ggtag 25
<210> 430
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 430
ggaaacgttc atggcaatta ttttc 25
<210> 431
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 431
cacccctttg tcatcttcgt tc 22
<210> 432
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 432
cacttccctc acaactgatg c 21
<210> 433
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 433
tgaattcact ccttagacct caatg 25
<210> 434
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 434
tctagtgcgt tcttacttct cac 23
<210> 435
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 435
tgaagtagat caaattggcc gg 22
<210> 436
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 436
ctatctccga tataggggta aagac 25
<210> 437
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 437
aaccatgcac tgggtagttc ta 22
<210> 438
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 438
gacggaagaa gaacagttag gg 22
<210> 439
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 439
gagcccctac tccgtgatcg 20
<210> 440
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 440
tctatggctt tgaatgataa gcagc 25
<210> 441
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 441
gcagcaacag cagatggagc 20
<210> 442
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 442
cgtggactat ggaggtcgac 20
<210> 443
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 443
tgagcaatac atttctgttg tagc 24
<210> 444
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 444
gctacctttc ttagctggga tcag 24
<210> 445
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 445
acgcaggtgc agaccaag 18
<210> 446
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 446
cgtccttgtc tgtttaaagc tct 23
<210> 447
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 447
tggccaagag tccagacatc 20
<210> 448
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 448
cctcaccact tgcaacataa tgc 23
<210> 449
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 449
tgaaataaat gaccctttct cctcg 25
<210> 450
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 450
gcattgctgg gtcatcattc g 21
<210> 451
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 451
ttctgtcaat atctttcttc agctg 25
<210> 452
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 452
accttcttct tcctccaatc cac 23
<210> 453
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 453
cgcatttggg atcaggttcc 20
<210> 454
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 454
aaaaggtcat cacaaaccaa aaac 24
<210> 455
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 455
tccaatgaat acaggatggt cc 22
<210> 456
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 456
ccaagtatct aattagttca gcctg 25
<210> 457
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 457
ggactcaaga gaatcatcat tgatc 25
<210> 458
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 458
tgacgctgtt aatcaattgc tcg 23
<210> 459
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 459
agtaagtcca gttccttcag tct 23
<210> 460
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 460
aattttggtg ccactccatt g 21
<210> 461
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 461
tgtcctaagt gctcatcatc cc 22
<210> 462
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 462
ttctaacaat ctcatcaacg acatc 25
<210> 463
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 463
agctgtggtc atattatgca atgtc 25
<210> 464
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 464
gaggtttctc taatgcacaa gcaca 25
<210> 465
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 465
tgcccctaac cagtaacttg c 21
<210> 466
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 466
ggctggcttt ttatatccaa ggc 23
<210> 467
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 467
cctagctcgg tttccagcg 19
<210> 468
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 468
cgaggattag tggcttttga acg 23
<210> 469
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 469
taaggcttca tcctcctggc 20
<210> 470
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 470
gaatctcggg ctcaaagtca ttt 23
<210> 471
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 471
ccagagtcaa cgatacctac gc 22
<210> 472
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 472
ttcctcttca tcccgttgat c 21
<210> 473
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 473
caaactgata tacactcacc tcaag 25
<210> 474
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 474
cgcgactgtc ggctgacg 18
<210> 475
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 475
tgcactacta gatggtggca tag 23
<210> 476
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 476
agttcacctt ctctacagtt tccc 24
<210> 477
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 477
cgtccgcaga ccttgaagtt atc 23
<210> 478
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 478
cagccacagc catgctcg 18
<210> 479
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 479
agtagggtac tcattccata attcc 25
<210> 480
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 480
cagagtccag atggctatac tgtc 24
<210> 481
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 481
acgtgtattg gagcgcttca 20
<210> 482
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 482
cactggctgc tgcagaaaat 20
<210> 483
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 483
cttctttttc tcatgatatc gtgga 25
<210> 484
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 484
agacctctat accggagcct 20
<210> 485
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 485
ccgttgccag tattgtaggt atttt 25
<210> 486
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 486
tctagggttg gagttaattc cct 23
<210> 487
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 487
tcgacacccc tgccaatttg 20
<210> 488
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 488
tcctgaactt gaaatggatg act 23
<210> 489
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 489
actcaaccac ctatctattc gtt 23
<210> 490
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 490
tgccatcctc ctacaagacc 20
<210> 491
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 491
ccccgaaatt tgcagccaaa 20
<210> 492
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 492
cacaatgtaa gagaaaggca gagt 24
<210> 493
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 493
aaccacttcc ccatttatga taga 24
<210> 494
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 494
ctactctgat ttgaagaaac tgtca 25
<210> 495
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 495
caacaatatt ggattcagtt gaacc 25
<210> 496
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 496
cgatccactt tctgatacac ttttt 25
<210> 497
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 497
ccccttgtcc ttcgtcatcc 20
<210> 498
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 498
caaatgcgac gacaacaact tt 22
<210> 499
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 499
tcacggggaa gaagaccagg 20
<210> 500
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 500
cgagcaacag ctatttcctc g 21
<210> 501
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 501
tatcttgagc atggtcagcc ttac 24
<210> 502
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 502
cctcttctag catccctgac at 22
<210> 503
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 503
cggagtgtta cagaccaagt ga 22
<210> 504
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 504
tgctgcagat tctgaatttg gg 22
<210> 505
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 505
actcatcatc atcatcgtca tcg 23
<210> 506
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 506
tgatttcagg tcaatatgtc tggt 24
<210> 507
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 507
tccgagatgc aactcttagc c 21
<210> 508
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 508
tgacgctgtt ccaggtgtac 20
<210> 509
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 509
accgaactgt ctttttatcc cg 22
<210> 510
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 510
tgcaagaatc tgttaatgca acatt 25
<210> 511
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 511
ccaccatgac cgagaataag aga 23
<210> 512
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 512
acttgactag acctccgttc tct 23
<210> 513
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 513
ttctctgttc ctctaaaccc tg 22
<210> 514
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 514
tctcacagtg ggttcccgt 19
<210> 515
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 515
cccaaggagg caagagcaac 20
<210> 516
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 516
tttgggtaca gggaagagcc 20
<210> 517
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 517
gcaaacgaaa aacaaagcgg ata 23
<210> 518
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 518
gctgtaactt taattgcatt tcctg 25
<210> 519
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 519
gatttttcgt gtgccggcag 20
<210> 520
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 520
cacctcgatg tgtaacagga aga 23
<210> 521
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 521
gccagagtat ttcttcaaat cttca 25
<210> 522
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 522
acagaagaag ctgcaatcac att 23
<210> 523
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 523
agaagtagaa gaagaagagg aggag 25
<210> 524
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 524
aaagagtttg gtcatgatca agag 24
<210> 525
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 525
cccttctctc cctggaatgt tg 22
<210> 526
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 526
agcttcgggg cctacatagg 20
<210> 527
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 527
gtgcttccca aagatatgaa atgc 24
<210> 528
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 528
tgccaagact gactacaact c 21
<210> 529
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 529
cagcaatggt agagctcaag c 21
<210> 530
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 530
cgaggccaca gacatagttt g 21
<210> 531
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 531
ggaacacgaa gaaggcctga 20
<210> 532
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 532
tctccaaatc cttaacggac ct 22
<210> 533
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 533
atgatctaaa ccctacatca cctt 24
<210> 534
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 534
gggctgagac caggagagt 19
<210> 535
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 535
ctcaagtcta cctaggacta agct 24
<210> 536
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 536
tctaaggaca tccaacactg cc 22
<210> 537
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 537
agcggcaaca tgtccagaat 20
<210> 538
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 538
gcttccagaa ctagtgtaga tactt 25
<210> 539
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 539
ggtcaatggt cgttgtttct ca 22
<210> 540
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 540
ccaaatctgt ggcccgcata 20
<210> 541
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 541
gtctatttcg taaatcttct gaccg 25
<210> 542
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 542
tcagttgaac ccgtaagcag t 21
<210> 543
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 543
aataccaggt gcactcgcca 20
<210> 544
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 544
cacttgacac ttctttcaga gattt 25
<210> 545
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 545
atggtgagga cgtgcacttc 20
<210> 546
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 546
cctggtgtca agctgcacta 20
<210> 547
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 547
acggcatgag agatacttaa acga 24
<210> 548
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 548
ccattactca catttccaac tggg 24
<210> 549
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 549
ctttccgcca cctcctccg 19
<210> 550
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 550
ataaaaggac atttcttcag gttgt 25
<210> 551
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 551
ttagaatcct tgggcataca gat 23
<210> 552
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 552
accctttgaa tgatgtcagc t 21
<210> 553
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 553
tgaagtgaca tctgatgacg aa 22
<210> 554
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 554
aggttgcaca tttttaccca aattc 25
<210> 555
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 555
tgcttatcca agattctcta gcata 25
<210> 556
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 556
cgatcctcct acgatgagca c 21
<210> 557
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 557
aatctggtgc aaccgcttct 20
<210> 558
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 558
taggagagac gtgccatggt 20
<210> 559
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 559
tgtgtgagaa aacatattgt ggca 24
<210> 560
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 560
acttatttac tcatgccgtg ttg 23
<210> 561
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 561
tggctcacta tctctagcaa tg 22
<210> 562
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 562
ggtatcactc ctttcaagtt tcaaa 25
<210> 563
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 563
agcaatctga atccatgtaa atgag 25
<210> 564
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 564
aggtgaagag tcctcctcag a 21
<210> 565
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 565
aggtaaaccg agtaagatgg ga 22
<210> 566
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 566
gcacaaggaa catgccttaa tcat 24
<210> 567
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 567
tcattcgtag agggctggga 20
<210> 568
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 568
tggagccaag catcatcaag ct 22
<210> 569
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 569
tggaattttc tcaggggcct 20
<210> 570
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 570
aacacagagg cattggcaag 20
<210> 571
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 571
agatgtccta cagtggtttc ac 22
<210> 572
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 572
gcccaggcaa tgtatgaaag 20
<210> 573
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 573
cgggtctcta ttaggctttt tcttc 25
<210> 574
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 574
caacctccat tctcctcttc tatat 25
<210> 575
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 575
tctgaagact cgataaaagc acttg 25
<210> 576
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 576
tgcttcctca acatctacat acg 23
<210> 577
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 577
acccagtatt tccgacacga 20
<210> 578
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 578
caaatattcc agtgtcgaga tgtta 25
<210> 579
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 579
ctttcatcgt tcacacaccg t 21
<210> 580
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 580
ttgctgctgt ccaaaagtct 20
<210> 581
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 581
gctgaagaaa accaagccca c 21
<210> 582
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 582
tcctgaactt tggcagttgt g 21
<210> 583
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 583
aatatcgaaa cccggacccc 20
<210> 584
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 584
tgcgacaaaa agatattatg aaccg 25
<210> 585
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 585
tggtttcaat tggttcaccc ac 22
<210> 586
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 586
ctacggctga gggaagtgga 20
<210> 587
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 587
cgaggggttg aacggatcat 20
<210> 588
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 588
tcttcaccca ccttttctat gtta 24
<210> 589
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 589
ggagcaatta gtggctctgc 20
<210> 590
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 590
aacagcaagt gcaaagatgg 20
<210> 591
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 591
cgaaggctgg gtcgtgaa 18
<210> 592
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 592
cagcaacgtt gtgtccacg 19
<210> 593
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 593
gaagcatccg cacaaccaaa 20
<210> 594
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 594
agaactacct aacccaccta ca 22
<210> 595
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 595
tggaacaaag ttagacctaa aggga 25
<210> 596
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 596
cctcctcacc cttacgtttg tt 22
<210> 597
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 597
ggcacaactt ttcacgccat 20
<210> 598
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 598
atgctagaga tagcggtgcc 20
<210> 599
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 599
agcaagaaga ggaaacagcc t 21
<210> 600
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 600
aaagaacgcg cttactttga at 22
<210> 601
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 601
agagacaaac catggtcttt tgg 23
<210> 602
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 602
gggcactaaa tgtcacacac a 21
<210> 603
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 603
tgagttgtat aacgctaggg gc 22
<210> 604
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 604
actaaaatta aaagcgcagt gga 23
<210> 605
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 605
acgagcatca gattttgttc ac 22
<210> 606
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 606
tatggtgtac cgtgtctcct tcatt 25
<210> 607
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 607
agtagcataa aatttaccag acgc 24
<210> 608
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 608
cgggggatag caaagaatca c 21
<210> 609
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 609
agccaagttg gacagtcgac 20
<210> 610
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 610
caacaattcc tgctaagaaa gagat 25
<210> 611
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 611
tgagacgtgt caatccagat aac 23
<210> 612
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 612
ttcggctcga gagaatgagt 20
<210> 613
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 613
tcgaatcctg tcgttgcgag 20
<210> 614
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 614
actttccggt gctaacgatg 20
<210> 615
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 615
gctatgtccg gttgccaca 19
<210> 616
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 616
cgaggatcga gatcatatac acat 24
<210> 617
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 617
ccatgtgttg tttcactccc tta 23
<210> 618
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 618
cctttagacc tcctcatgct ca 22
<210> 619
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 619
caagatctca caactcctct aattc 25
<210> 620
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 620
accatttggc atgtactcgt at 22
<210> 621
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 621
tgctgtttta gtgtttgatg gatt 24
<210> 622
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 622
cctagatgga tgaggttcta tactg 25
<210> 623
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 623
gtgagagttt ccacttatcc gact 24
<210> 624
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 624
agtatgtgga gttctctagc aga 23
<210> 625
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 625
tggaataagg tcctacccct t 21
<210> 626
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 626
agtaccagat tcattgccca tat 23
<210> 627
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 627
agtgtcggtg gaggctcg 18
<210> 628
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 628
tgatttgtat agtcacttga gaagc 25
<210> 629
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 629
tctttgtctt tttctagcaa actga 25
<210> 630
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 630
agcgacaaag cacctgaaaa g 21
<210> 631
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 631
acaattccac ataaatagca acctt 25
<210> 632
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 632
agagatgatc tatcacctta acccc 25
<210> 633
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 633
gcctccagtt ctcttcggag 20
<210> 634
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 634
cgactacctc cacacgatcg 20
<210> 635
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 635
gaacgattcg agcgtacttt c 21
<210> 636
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 636
tcccaggagc ctcaagtgta 20
<210> 637
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 637
ccccgacggt gagaaagac 19
<210> 638
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 638
gcactccacc ccatagtcaa 20
<210> 639
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 639
ccgatttggc tgacttatgt atttt 25
<210> 640
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 640
acgtacatgt caggcccaaa 20
<210> 641
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 641
aattctacca ccaaaactgc ac 22
<210> 642
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 642
gatcacttac agttcaaaaa gcttg 25
<210> 643
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 643
gagctgaatc aggagaggga g 21
<210> 644
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 644
aatgcaaaac aagaaacctg tttta 25
<210> 645
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 645
ttcaacagct ccttcacgcc 20
<210> 646
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 646
cacataaact ggagcggagg ta 22
<210> 647
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 647
actgtgtggg attgccttct 20
<210> 648
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 648
ggcaaccctt cccctttc 18
<210> 649
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 649
agaaggttct gaggttggcg 20
<210> 650
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 650
accaagtgct ataccaaatg ga 22
<210> 651
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 651
tcttcgatga taactgggta tgttc 25
<210> 652
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 652
tgccaaggac gtacatccca 20
<210> 653
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 653
ggcctggaga caacgtgttt 20
<210> 654
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 654
ggtttagcac tgtcttggca g 21
<210> 655
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 655
agtagcggta catgatacat ttgt 24
<210> 656
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 656
cacccgactg cccttttgtt 20
<210> 657
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 657
tgaagagacg ttaagggcca 20
<210> 658
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 658
tggcaaaagt ttcttcagtt gct 23
<210> 659
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 659
attccgacga cttctccgct 20
<210> 660
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 660
gaacgttcca agcaagcctg 20
<210> 661
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 661
tctcgaaaga tgaagcctgt gta 23
<210> 662
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 662
tgataaccta cctctagctg aatga 25
<210> 663
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 663
tcaccaaaca gctaaaagag gtaaa 25
<210> 664
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 664
gatgcaacca tatctttgaa gtgac 25
<210> 665
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 665
acactaccct tctcagaccc t 21
<210> 666
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 666
ccaccatggc caactctcc 19
<210> 667
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 667
agagctcaaa gccatagtct cc 22
<210> 668
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 668
actcgaatcg actgatcaat catct 25
<210> 669
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 669
cagagttttg gattccttgt tttct 25
<210> 670
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 670
cgagtttgag aataatggct acca 24
<210> 671
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 671
aagggaagga acacaacgca 20
<210> 672
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 672
aggtatgcaa gaacagatga actt 24
<210> 673
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 673
cgtccacgtt aacagaggat ttg 23
<210> 674
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 674
acacactcca attccagatg tttt 24
<210> 675
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 675
caacctgcag ccatgttgaa t 21
<210> 676
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 676
cgcgttatgc accacaacaa 20
<210> 677
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 677
ggtaggcatt gggtgtgtct 20
<210> 678
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 678
tggtactacc gatgacaacg ttt 23
<210> 679
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 679
tgctttcggg ccatctcttt 20
<210> 680
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 680
ctgccatttt ccgctttctt g 21
<210> 681
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 681
accaacctaa cctataatgt tcctc 25
<210> 682
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 682
tccataaaag aaaacgttgt gtgt 24
<210> 683
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 683
tcatccgttg caccacatcc 20
<210> 684
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 684
gatgtgaatg tctttggggc a 21
<210> 685
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 685
ctcgcagatg tgaaccgttg 20
<210> 686
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 686
catgtggatg ctcctggctt 20
<210> 687
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 687
acgttcttga agtgctactt tacta 25
<210> 688
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 688
tgaaggtagg gtgggaccaa 20
<210> 689
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 689
gtttccagga gaccctcagc 20
<210> 690
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 690
acccagatgt gcatatttta cagag 25
<210> 691
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 691
cgatccaaac atatgggata attga 25
<210> 692
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 692
acaagagaaa gagtgagagc gt 22
<210> 693
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 693
ttccaaagtt cgaccgcaga 20
<210> 694
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 694
cctgaaatga atgccctttg attta 25
<210> 695
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 695
gtcatgaaga tagggccatc acta 24
<210> 696
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 696
agcacagttc gaactacaac ct 22
<210> 697
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 697
agtatcacca actcaatatc tggag 25
<210> 698
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 698
agaatgatta aactccgtac agca 24
<210> 699
<211> 19
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 699
ggcaacgact tggggaagg 19
<210> 700
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 700
tctgggttgt tgttgttgtt gt 22
<210> 701
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 701
tccgggaata ccttcaagcc 20
<210> 702
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 702
aggtttggac aatgcaattg g 21
<210> 703
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 703
tccactctca gctttactcc a 21
<210> 704
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 704
tcatgttgca acgaatcctt gtt 23
<210> 705
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 705
ttggctagcg tcggacttac 20
<210> 706
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 706
gggaagagcg atcttctgga 20
<210> 707
<211> 22
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 707
actatcatct gtgcctttgg tg 22
<210> 708
<211> 18
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 708
aatccccctt cggctccc 18
<210> 709
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 709
tgcccacata ttcactagtg ctata 25
<210> 710
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 710
attccaccgt agccgtagtt 20
<210> 711
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 711
tcagcctctg atatctctcc a 21
<210> 712
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 712
agagccgtac tagtggagtg a 21
<210> 713
<211> 23
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 713
ggaggaaagc cgaaaatatc aac 23
<210> 714
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 714
gggtagagga ggagtagagc a 21
<210> 715
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 715
gggacctctt tttactttaa gacca 25
<210> 716
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 716
cgtcatcccc tgctattcag t 21
<210> 717
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 717
agccaattcc tctaaaacaa ctaaa 25
<210> 718
<211> 24
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 718
cggttggatg agcgattatt taga 24
<210> 719
<211> 20
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 719
ggttgcttga cccctacctc 20
<210> 720
<211> 25
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 720
gttctagttt tctctttggt acagg 25
<210> 721
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 721
gaaggtgacc aagttcatgc t 21
<210> 722
<211> 21
<212> DNA
<213> Artificial sequence (Artificial sequence)
<400> 722
gaaggtcgga gtcaacggat t 21

Claims (10)

1. The application of the reagent for detecting the genotypes of 240 SNP loci in the tobacco genome in any one of the following steps:
(1) Preparing a kit for detecting the genotype of the SNP locus of the tobacco;
(2) Genotyping a tobacco germplasm resource or genetic population;
(3) Constructing a DNA fingerprint of tobacco germplasm resources or varieties;
(4) Detecting the purity or the authenticity of the tobacco variety;
the type of tobacco is selected from flue-cured tobacco, sun-cured tobacco, burley tobacco, cigar tobacco, aromatic tobacco and medicinal tobacco;
the physical positions of the 240 SNP loci are determined based on the comparison of the whole genome sequence of the tobacco variety Honghuadajinyuan, and the version number of the whole genome sequence of the tobacco variety Honghuadajinyuan is V1.2; the 240 SNP sites are Ts001-Ts240;
ts001 is located on chromosome 1 at position 8208363 with the deoxynucleotide being T or C;
ts002 is located at 33067181 on chromosome 1, and its deoxynucleotide is G or A;
ts003 is located on chromosome 1 at position 57320321 with the deoxynucleotide G or a;
ts004 is located at position 81583864 of chromosome 1, and the deoxynucleotide of the Ts is A or C;
ts005 is located at 106322334 th on chromosome 1, and its deoxynucleotide is G or A;
ts006 is located on chromosome 1 at position 125569732 with the deoxynucleotide being T or C;
ts007 is located at 146339969 th position on chromosome 1, and the deoxynucleotide of the Ts is A or G;
ts008 is located at 168939497 th position on chromosome 1, and the deoxynucleotide is A or G;
ts009 is located at position 192328872 on chromosome 1, and its deoxynucleotide is A or G;
ts010 is located at 218318116 of chromosome 1, and its deoxynucleotide is C or T;
ts011 is located at 450633 th position on chromosome 2, and deoxynucleotide of the Ts is C or T;
ts012 is located at position 27809195 on chromosome 2, and its deoxynucleotide is A or T;
ts013 is located at 44225412 on chromosome 2, and its deoxynucleotide is C or T;
ts014 is located at position 62278727 on chromosome 2, and its deoxynucleotide is C or T;
ts015 is located at position 83667001 on chromosome 2, its deoxynucleotide is A or G;
ts016 is located on chromosome 2 at position 113116215, and its deoxynucleotide is G or A;
ts017 is positioned at 137528012 th position on chromosome 2, and the deoxynucleotide is G or A;
ts018 is located on chromosome 2 at position 169132158 with T or C deoxynucleotides;
ts019 is located at position 185483126 on chromosome 2 with the deoxynucleotide being G or A;
ts020 is located on chromosome 2 at position 208449625, and its deoxynucleotide is C or T;
ts021 is located on 70093 th position on chromosome 3, and the deoxynucleotide is T or C;
ts022 is positioned at the 27066915 th site on the chromosome 3, and the deoxynucleotide of the Ts is A or G;
ts023 is located at position 54763744 on chromosome 3, and its deoxynucleotide is G or A;
ts024 is located at position 70866040 on chromosome 3, and its deoxynucleotide is A or G;
ts025 is located at position 91669016 on chromosome 3, and its deoxynucleotide is C or T;
ts026 is located on chromosome 3 at position 116024385, and its deoxynucleotide is G or A;
ts027 is located on chromosome 3 at position 136069307, with the deoxynucleotide being T or G;
ts028 is located on chromosome 3 at position 156967681 with a deoxynucleotide of G or A;
ts029 is located on chromosome 3 at position 173064283, and the deoxynucleotide is A or G;
ts030 is located on chromosome 3 at position 198017426, and its deoxynucleotide is T or C;
ts031 is located at position 3899155 on chromosome 4 with deoxynucleotide G or A;
ts032 is located at position 15517251 on chromosome 4, and the deoxynucleotide is C or T;
ts033 is located at position 26648428 on chromosome 4, and its deoxynucleotide is C or G;
ts034 is located on chromosome 4 at position 46186327 and its deoxynucleotide is C or T;
ts035 is 63336872 on chromosome 4 with deoxynucleotide C or G;
ts036 is located at position 83168757 on chromosome 4, and its deoxynucleotide is A or G;
ts037 is located at position 94688224 on chromosome 4, and its deoxynucleotide is C or T;
ts038 is located on chromosome 4 at position 105682078, and its deoxynucleotide is C or T;
ts039 is located at position 124010732 on chromosome 4 with the deoxynucleotide being C or G;
ts040 is located at position 136418400 on chromosome 4, and its deoxynucleotide is C or G;
ts041 is located at position 818540 on chromosome 5, and its deoxynucleotide is G or C;
ts042 is located on chromosome 5 at position 13565159, and the deoxynucleotide is G or T;
ts043 is located on chromosome 5 at position 27317490, and the deoxynucleotide is C or T;
ts044 is located on chromosome 5 at position 42181447, and the deoxynucleotide is G or A;
ts045 is located at 67747378 th position on chromosome 5, and the deoxynucleotide is A or T;
ts046 is located at 80270025 on chromosome 5, and the deoxynucleotide is G or T;
ts047 is located on chromosome 5 at position 88119523, and its deoxynucleotide is A or G;
ts048 is located on chromosome 5 at position 98029128, and its deoxynucleotide is T or A;
ts049 is located on chromosome 5 at position 127826992, and its deoxynucleotide is A or T;
ts050 is located at position 133049305 on chromosome 5, and the deoxynucleotide of the Ts is A or T;
ts051 is located at position 13578189 on chromosome 6, and the deoxynucleotide is C or T;
ts052 is located on chromosome 6 at position 25353358, and its deoxynucleotide is G or A;
ts053 is located on chromosome 6 at position 4424521, and its deoxynucleotide is T or C;
ts054 is located on chromosome 6 at position 58884603 with the deoxynucleotide G or a;
ts055 is located at 72393591 of chromosome 6, and its deoxynucleotide is A or G;
ts056 is located at position 84232195 on chromosome 6, and its deoxynucleotide is C or T;
ts057 is located at 95593235 on chromosome 6, and its deoxynucleotide is A or C;
ts058 is located on chromosome 6 at position 115033653, and its deoxynucleotide is T or A;
ts059 is located on chromosome 6 at position 125125125703, and its deoxynucleotide is A or G;
ts060 is located at the 141918758 th site on chromosome 6, and the deoxynucleotide is C or T;
ts061 is located on chromosome 7 at position 414141224 with deoxynucleotides C or T;
ts062 is located at position 14082999 on chromosome 7, and the deoxynucleotide is C or A;
ts063 is located at 22079279 on chromosome 7, and its deoxynucleotide is C or T;
ts064 is located at position 36706020 on chromosome 7, and the deoxynucleotide is G or T;
ts065 is located at 51446290 position on chromosome 7, and the deoxynucleotide is T or C;
ts066 is located at position 64707494 on chromosome 7, and the deoxynucleotide of the Ts is T or A;
ts067 is located at 79594403 th position on chromosome 7, and the deoxynucleotide of the Ts067 is G or A;
ts068 is located at position 100880287 on chromosome 7 with the deoxynucleotide being C or G;
ts069 is located at position 116533979 on chromosome 7, and its deoxynucleotide is A or C;
ts070 is located on chromosome 7 at position 134235128, and the deoxynucleotide is A or T;
ts071 is located at 3230578 th position on chromosome 8, and its deoxynucleotide is G or A;
ts072 is located on chromosome 8 at position 14779614 with deoxynucleotides C or T;
ts073 is located on chromosome 8 at position 34418092, and its deoxynucleotide is C or A;
ts074 is located on chromosome 8 at position 47428516, and its deoxynucleotide is G or A;
ts075 is located on chromosome 8 at position 64510378, and its deoxynucleotide is G or T;
ts076 is located at position 77171808 on chromosome 8, and its deoxynucleotide is A or C;
ts077 is located at 86364114 th on chromosome 8, and the deoxynucleotide is T or C;
ts078 is located on chromosome 8 at position 103332416, and its deoxynucleotide is A or G;
ts079 is located on chromosome 8 at position 119385289, and its deoxynucleotide is C or G;
ts080 is located at 139629563 on chromosome 8, and the deoxynucleotide thereof is C or T;
ts081 is located at position 10705780 on chromosome 9, and its deoxynucleotide is A or T;
ts082 is located at position 25510126 on chromosome 9, and its deoxynucleotide is A or G;
ts083 is located at position 36791110 on chromosome 9, and its deoxynucleotide is C or T;
ts084 is located on chromosome 9 at position 52279521, and its deoxynucleotide is C or A;
ts085 is located at position 66389919 on chromosome 9, and its deoxynucleotide is A or G;
ts086 is located on chromosome 9 at position 76596627 with deoxynucleotide T or C;
ts087 is located at position 88782369 on chromosome 9, and its deoxynucleotide is A or G;
ts088 is located at position 101792480 on chromosome 9, and the deoxynucleotide is A or G;
ts089 is located on chromosome 9 at position 114504225, and its deoxynucleotide is A or G;
ts090 is located at position 125921087 on chromosome 9, and the deoxynucleotide is C or T;
ts091 is located at position 8333061 on chromosome 10, and its deoxynucleotide is T or G;
ts092 is located on chromosome 10 at position 19900537 with a deoxynucleotide that is T or C;
ts093 is located on chromosome 10 at position 30838441, and its deoxynucleotide is A or G;
ts094 is located on chromosome 10 at position 48459980, and its deoxynucleotide is A or T;
ts095 is located on chromosome 10 at position 60720808, and its deoxynucleotide is A or G;
ts096 is located on chromosome 10 at position 81556459, and its deoxynucleotide is G or C;
ts097 is located on chromosome 10 at position 91772677, and its deoxynucleotide is G or A;
ts098 is located on chromosome 10 at position 104792976 with a deoxynucleotide as G or A;
ts099 is located on chromosome 10 at position 115447057 with a deoxynucleotide as G or A;
ts100 is located on chromosome 10 at position 127459715, and the deoxynucleotide of the Ts is A or G;
ts101 is located on chromosome 11 at position 1849343, and its deoxynucleotide is C or T;
ts102 is located at position 9349425 on chromosome 11, and its deoxynucleotide is A or G;
ts103 is located at 21455270 th on chromosome 11, and the deoxynucleotide is C or T;
ts104 is positioned at 26997488 th position on chromosome 11, and the deoxynucleotide of the Ts is C or T;
ts105 is located at position 39424005 on chromosome 11, and the deoxynucleotide of the Ts is T or G;
ts106 is located at the position 54261766 on chromosome 11, and the deoxynucleotide of the Ts is G or T;
ts107 is located on chromosome 11 at position 71261702 with the deoxynucleotide being T or G;
ts108 is located on chromosome 11 at position 89921212, and its deoxynucleotide is T or A;
ts109 is located on chromosome 11 at position 108191371, and its deoxynucleotide is G or A;
ts110 is located on chromosome 11 at position 125710919, and its deoxynucleotide is C or T;
ts111 is located on chromosome 12 at position 6346289, and its deoxynucleotide is G or A;
ts112 is located on chromosome 12 at position 19411089, and its deoxynucleotide is G or C;
ts113 is located at position 35572511 on chromosome 12, and its deoxynucleotide is C or A;
ts114 is located on chromosome 12 at position 50563895, and its deoxynucleotide is C or A;
ts115 is located at position 64428830 on chromosome 12, and the deoxynucleotide of the Ts is A or G;
ts116 is located at 72201030 on chromosome 12, and the deoxynucleotide is C or T;
ts117 is located on chromosome 12 at position 84715140 with deoxynucleotide A or G;
ts118 is located on chromosome 12 at position 96783131 with the deoxynucleotide being C or T;
ts119 is located on chromosome 12 at position 112836712, and its deoxynucleotide is C or A;
ts120 is located on chromosome 12 at position 120317419, and its deoxynucleotide is C or T;
ts121 is located on chromosome 13 at position 2084818, and its deoxynucleotide is A or G;
ts122 is located on chromosome 13 at position 15919260, and its deoxynucleotide is A or T;
ts123 is located at position 29366753 on chromosome 13, and the deoxynucleotide is G or T;
ts124 is located on the No.13 chromosome at the 42405167 position, and the deoxynucleotide is G or A;
ts125 is located on chromosome 13 at position 50730421 with a deoxynucleotide T or C;
ts126 is located on chromosome 13 at position 60261050, and the deoxynucleotide is G or A;
ts127 is located on chromosome 13 at position 71762485, and its deoxynucleotide is C or A;
ts128 is located on chromosome 13 at position 86588177, and its deoxynucleotide is C or T;
ts129 is located at position 100565007 on chromosome 13, and the deoxynucleotide of the Ts is A or G;
ts130 is located on chromosome 13 at position 111060065, and the deoxynucleotide is T or G;
ts131 is located on chromosome 14 at position 761393, and its deoxynucleotide is G or A;
ts132 is located on chromosome 14 at position 10247910 with the deoxynucleotide being C or T;
ts133 is located at position 25145170 on chromosome 14 with the deoxynucleotide being G or A;
ts134 is located on chromosome 14 at position 33808789, and its deoxynucleotide is A or G;
ts135 is located on chromosome 14 at position 43276550, and its deoxynucleotide is G or A;
ts136 is located on chromosome 14 at position 54717027 with deoxynucleotides T or C;
ts137 is located at position 64685001 on chromosome 14, and its deoxynucleotide is G or T;
ts138 is located on chromosome 14 at position 76697305, and the deoxynucleotide of the Ts is C or T;
ts139 is located on chromosome 14 at position 89419411, and its deoxynucleotide is A or T;
ts140 is located on chromosome 14 at position 105075736 with the deoxynucleotide a or G;
ts141 is located on chromosome 15 at position 151458 with the deoxynucleotide being T or C;
ts142 is located at position 9949166 on chromosome 15, and its deoxynucleotide is G or A;
ts143 is located on chromosome 15 at position 20982657, and its deoxynucleotide is A or G;
ts144 is located on chromosome 15 at position 31321347 with the deoxynucleotide being G or T;
ts145 is located on chromosome 15 at position 43232491, and its deoxynucleotide is A or C;
ts146 is located on the No.15 chromosome at position 49438846, and the deoxynucleotide of the Ts is G or C;
ts147 is located at 65072165 on chromosome 15, and the deoxynucleotide is C or T;
ts148 is located on chromosome 15 at position 76351185, and its deoxynucleotide is A or C;
ts149 is located on chromosome 15 at position 89189533 with the deoxynucleotide being G or A;
ts150 is located on chromosome 15 at position 104194974 with the deoxynucleotide being T or G;
ts151 is located on chromosome 16 at position 1110051, and its deoxynucleotide is T or C;
ts152 is located at 10087681 position on chromosome 16, and the deoxynucleotide of the Ts is C or T;
ts153 is located at 17842746 on chromosome 16, and the deoxynucleotide is C or T;
ts154 is located at position 26067906 on chromosome 16, and its deoxynucleotide is A or C;
ts155 is located on chromosome 16 at position 34995199, and its deoxynucleotide is G or A;
ts156 is located on chromosome 16 at position 46203805 with the deoxynucleotide being T or C;
ts157 is located on chromosome 16 at position 52542790 with the deoxynucleotide being T or C;
ts158 is located on chromosome 16 at position 62428942, and its deoxynucleotide is C or G;
ts159 is located on chromosome 16 at position 79498876 with the deoxynucleotide being C or T;
ts160 is located on chromosome 16 at position 82884529 with the deoxynucleotide C or a;
ts161 is located on chromosome 17 at position 3824750 with the deoxynucleotide being A or G;
ts162 is located on chromosome 17 at position 13454308 with the deoxynucleotide C or T;
ts163 is located on chromosome 17 at position 23361027 with a T or C deoxynucleotide;
ts164 is located on chromosome 17 at position 30960072, and its deoxynucleotide is T or A;
ts165 is located on chromosome 17 at position 3985348, and its deoxynucleotide is G or A;
ts166 is located on chromosome 17 at position 49241377, and its deoxynucleotide is G or A;
ts167 is located on chromosome 17 at position 56588921 with deoxynucleotides a or G;
ts168 is located on chromosome 17 at position 68788574 with the deoxynucleotide being T or a;
ts169 is located on chromosome 17 at position 76261437, and its deoxynucleotide is A or T;
ts170 is located on chromosome 17 at position 86191077, and its deoxynucleotide is C or T;
ts171 is located on chromosome 18 at position 10666388 with the deoxynucleotide being T or G;
ts172 is located on chromosome 18 at position 23470800 with the deoxynucleotide a or G;
ts173 is located on chromosome 18 at position 30405985 with a deoxynucleotide of C or T;
ts174 is located on chromosome 18 at position 37097793, and its deoxynucleotide is C or A;
ts175 is located on chromosome 18 at position 41051715 with the deoxynucleotide being T or A;
ts176 is located on chromosome 18 at position 49088497, and the deoxynucleotide is C or A;
ts177 is located on chromosome 18 at position 58652117, and its deoxynucleotide is G or T;
ts178 is located on chromosome 18 at position 69344463, and its deoxynucleotide is T or C;
ts179 is located at position 77733306 on chromosome 18, and its deoxynucleotide is T or A;
ts180 is located on chromosome 18 at position 84321700, and its deoxynucleotide is G or A;
ts181 is located at position 2073784 on chromosome 19 with the deoxynucleotide being T or A;
ts182 is located on chromosome 19 at position 7537621 with the deoxynucleotide being G or a;
ts183 is located on chromosome 19 at position 13153286, and its deoxynucleotide is C or T;
ts184 is located at 17723705 th on chromosome 19, and its deoxynucleotide is C or G;
ts185 is located on chromosome 19 at position 27550742, and its deoxynucleotide is T or C;
ts186 is located on chromosome 19 at position 32885827 with the deoxynucleotide being a or G;
ts187 is located at position 39636484 on chromosome 19, and its deoxynucleotide is G or A;
ts188 is located on chromosome 19 at position 44936247, and the deoxynucleotide is C or T;
ts189 is located at 57465608 on chromosome 19, and its deoxynucleotide is C or T;
ts190 is located on chromosome 19 at position 74639708, and its deoxynucleotide is C or T;
ts191 is located on chromosome 20 at position 4182297 with the deoxynucleotide being G or A;
ts192 is located at position 13966980 on chromosome 20, and its deoxynucleotide is G or A;
ts193 is located on chromosome 20 at position 19609796 with the deoxynucleotide being G or a;
ts194 is located at position 29237378 on chromosome 20, and the deoxynucleotide is A or G;
ts195 is located on chromosome 20 at position 36180655, and its deoxynucleotide is T or G;
ts196 is located on chromosome 20 at position 39861635, with the deoxynucleotide being G or C;
ts197 is located on chromosome 20 at position 52409493, and its deoxynucleotide is T or A;
ts198 is positioned at the 56616994 th site on the 20 th chromosome, and the deoxynucleotide is C or T;
ts199 is located on chromosome 20 at position 66112600 with the deoxynucleotide being G or T;
ts200 is located on chromosome 20 at position 76769655 with the deoxynucleotide being G or A;
ts201 is located at position 1120311 on chromosome 21, and its deoxynucleotide is A or G;
ts202 is located on chromosome 21 at position 8841119 with the deoxynucleotide being T or G;
ts203 is located at position 15008035 on chromosome 21, and the deoxynucleotide is G or A;
ts204 is located on chromosome 21 at position 21697573, with the deoxynucleotide being T or G;
ts205 is located on chromosome 21 at position 29569502, and its deoxynucleotide is C or T;
ts206 is located at position 35545304 on chromosome 21, and its deoxynucleotide is A or T;
ts207 is located on chromosome 21 at position 43960675, and its deoxynucleotide is C or T;
ts208 is located on chromosome 21 at position 52269430, and its deoxynucleotide is A or G;
ts209 is located at the 60583608 th position on the 21 st chromosome, and the deoxynucleotide of the Ts is A or G;
ts210 is located at 72870875 th position on chromosome 21, and the deoxynucleotide of the Ts is A or C;
ts211 is located on chromosome 22 at position 473170 with the deoxynucleotide being a or G;
ts212 is located on chromosome 22 at position 8509606 with the deoxynucleotide being T or C;
ts213 is located on chromosome 22 at position 12853488, and its deoxynucleotide is T or C;
ts214 is located at position 22117353 on chromosome 22, and its deoxynucleotide is G or A;
ts215 is located at position 26228906 on chromosome 22 with the deoxynucleotide being T or A;
ts216 is located on chromosome 22 at position 33899956, and its deoxynucleotide is C or T;
ts217 is located on chromosome 22 at position 41179872 with the deoxynucleotide being C or a;
ts218 is located on chromosome 22 at position 47452531 with the deoxynucleotide being C or G;
ts219 is located on chromosome 22 at position 58059052, and its deoxynucleotide is T or A;
ts220 is located at position 72943832 on chromosome 22, and its deoxynucleotide is A or G;
ts221 is located at position 79892 on chromosome 23 with the deoxynucleotide C or T;
ts222 is located on chromosome 23 at position 5555730, and its deoxynucleotide is C or T;
ts223 is located on chromosome 23 at position 13135791 with deoxynucleotides A or G;
ts224 is located on chromosome 23 at position 24118597 with the deoxynucleotide being A or T;
ts225 is located on chromosome 23 at position 34819441, and its deoxynucleotide is G or A;
ts226 is located on chromosome 23 at position 42069769, and its deoxynucleotide is T or C;
ts227 is located on chromosome 23 at position 46079860, with the deoxynucleotide being G or a;
ts228 is located on chromosome 23 at position 50972058 with the deoxynucleotide being a or G;
ts229 is located on chromosome 23 at position 58370965, with the deoxynucleotide being C or T;
ts230 is located at position 6624583 on chromosome 23, and its deoxynucleotide is A or T;
ts231 is located on chromosome 24 at position 5769442, and its deoxynucleotide is C or T;
ts232 is located on chromosome 24 at position 16375603, and its deoxynucleotide is T or C;
ts233 is located on chromosome 24 at position 19508580, and its deoxynucleotide is G or A;
ts234 is located on chromosome 24 at position 20893673, and its deoxynucleotide is T or G;
ts235 is located at position 27208986 on chromosome 24, and the deoxynucleotide is T or C;
ts236 is located on chromosome 24 at position 34358980, and its deoxynucleotide is C or T;
ts237 is located at position 44075839 on chromosome 24, and its deoxynucleotide is T or G;
ts238 is located on chromosome 24 at position 49853976, and its deoxynucleotide is A or G;
ts239 is located at position 58945156 on chromosome 24, and the deoxynucleotide of the Ts is T or G;
ts240 is located on chromosome 24 at position 63099098 with a deoxynucleotide as C or G.
2. A set of KASP primers for detecting the 240 SNP sites of claim 1;
the KASP primers corresponding to each SNP site to be detected respectively comprise two forward primers and one reverse primer; wherein two forward primers are respectively marked as a forward primer 1 and a forward primer 2; the 5 'end of the forward primer 1 is connected with a fluorescent label sequence, and the 5' end of the forward primer 2 is connected with another fluorescent label sequence;
the nucleotide sequence of a forward primer 1 for detecting Ts001 is shown as SEQ ID No.1, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.241, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 481;
the nucleotide sequence of the forward primer 1 for detecting Ts002 is shown as SEQ ID No.2, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.242, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 482;
the nucleotide sequence of a forward primer 1 for detecting Ts003 is shown as SEQ ID No.3, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.243, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 483;
the nucleotide sequence of the forward primer 1 for detecting Ts004 is shown as SEQ ID No.4, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.244, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 484;
the nucleotide sequence of the forward primer 1 for detecting Ts005 is shown as SEQ ID No.5, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.245, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 485;
the nucleotide sequence of a forward primer 1 for detecting Ts006 is shown as SEQ ID No.6, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.246, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 486;
the nucleotide sequence of the forward primer 1 for detecting Ts007 is shown as SEQ ID No.7, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.247, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 487;
the nucleotide sequence of a forward primer 1 for detecting Ts008 is shown as SEQ ID No.8, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.248, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 488;
the nucleotide sequence of the forward primer 1 for detecting Ts009 is shown as SEQ ID No.9, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.249, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 489;
the nucleotide sequence of the forward primer 1 for detecting Ts010 is shown as SEQ ID No.10, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.250, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 490;
the nucleotide sequence of a forward primer 1 for detecting Ts011 is shown in SEQ ID No.11, the nucleotide sequence of a forward primer 2 is shown in SEQ ID No.251, and the nucleotide sequence of a reverse primer is shown in SEQ ID No. 491;
the nucleotide sequence of a forward primer 1 for detecting Ts012 is shown as SEQ ID No.12, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.252, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 492;
the nucleotide sequence of a forward primer 1 for detecting Ts013 is shown in SEQ ID No.13, the nucleotide sequence of a forward primer 2 is shown in SEQ ID No.253, and the nucleotide sequence of a reverse primer is shown in SEQ ID No. 493;
the nucleotide sequence of a forward primer 1 for detecting Ts014 is shown as SEQ ID No.14, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.254, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 494;
the nucleotide sequence of a forward primer 1 for detecting Ts015 is shown as SEQ ID No.15, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.255, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 495;
the nucleotide sequence of a forward primer 1 for detecting Ts016 is shown as SEQ ID No.16, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.256, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 496;
the nucleotide sequence of the forward primer 1 for detecting Ts017 is shown as SEQ ID No.17, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.257, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 497;
the nucleotide sequence of a forward primer 1 for detecting Ts018 is shown as SEQ ID No.18, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.258, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 498;
the nucleotide sequence of the forward primer 1 for detecting Ts019 is shown as SEQ ID No.19, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.259, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 499;
the nucleotide sequence of the forward primer 1 for detecting Ts020 is shown in SEQ ID No.20, the nucleotide sequence of the forward primer 2 is shown in SEQ ID No.260, and the nucleotide sequence of the reverse primer is shown in SEQ ID No. 500;
the nucleotide sequence of the forward primer 1 for detecting Ts021 is shown as SEQ ID No.21, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.261, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 501;
the nucleotide sequence of a forward primer 1 for detecting Ts022 is shown as SEQ ID No.22, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.262, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 502;
the nucleotide sequence of the forward primer 1 for detecting Ts023 is shown as SEQ ID No.23, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.263, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 503;
the nucleotide sequence of a forward primer 1 for detecting Ts024 is shown as SEQ ID No.24, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.264, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 504;
the nucleotide sequence of the forward primer 1 for detecting Ts025 is shown as SEQ ID No.25, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.265, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 505;
the nucleotide sequence of the forward primer 1 for detecting Ts026 is shown as SEQ ID No.26, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.266, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 506;
the nucleotide sequence of the forward primer 1 for detecting Ts027 is shown in SEQ ID No.27, the nucleotide sequence of the forward primer 2 is shown in SEQ ID No.267, and the nucleotide sequence of the reverse primer is shown in SEQ ID No. 507;
the nucleotide sequence of the forward primer 1 for detecting Ts028 is shown as SEQ ID No.28, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.268, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 508;
the nucleotide sequence of a forward primer 1 for detecting Ts029 is shown as SEQ ID No.29, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.269, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 509;
the nucleotide sequence of a forward primer 1 for detecting Ts030 is shown as SEQ ID No.30, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.270, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 510;
the nucleotide sequence of the forward primer 1 for detecting Ts031 is shown as SEQ ID No.31, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.271, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 511;
the nucleotide sequence of the forward primer 1 for detecting Ts032 is shown as SEQ ID No.32, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.272, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 512;
the nucleotide sequence of a forward primer 1 for detecting Ts033 is shown as SEQ ID No.33, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.273, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 513;
the nucleotide sequence of a forward primer 1 for detecting Ts034 is shown as SEQ ID No.34, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.274, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 514;
the nucleotide sequence of the forward primer 1 for detecting Ts035 is shown as SEQ ID No.35, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.275, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 515;
the nucleotide sequence of the forward primer 1 for detecting Ts036 is shown as SEQ ID No.36, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.276, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 516;
the nucleotide sequence of a forward primer 1 for detecting Ts037 is shown as SEQ ID No.37, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.277, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 517;
the nucleotide sequence of a forward primer 1 for detecting Ts038 is shown as SEQ ID No.38, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.278, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 518;
the nucleotide sequence of a forward primer 1 for detecting Ts039 is shown as SEQ ID No.39, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.279, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 519;
the nucleotide sequence of the forward primer 1 for detecting Ts040 is shown as SEQ ID No.40, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.280, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 520;
the nucleotide sequence of the forward primer 1 for detecting the Ts041 is shown as SEQ ID No.41, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.281, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 521;
the nucleotide sequence of the forward primer 1 for detecting Ts042 is shown as SEQ ID No.42, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.282, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 522;
the nucleotide sequence of the forward primer 1 for detecting Ts043 is shown as SEQ ID No.43, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.283, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 523;
the nucleotide sequence of a forward primer 1 for detecting Ts044 is shown as SEQ ID No.44, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.284, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 524;
the nucleotide sequence of the forward primer 1 for detecting Ts045 is shown as SEQ ID No.45, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.285, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 525;
the nucleotide sequence of a forward primer 1 for detecting Ts046 is shown as SEQ ID No.46, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.286, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 526;
the nucleotide sequence of the forward primer 1 for detecting Ts047 is shown as SEQ ID No.47, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.287, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 527;
the nucleotide sequence of the forward primer 1 for detecting Ts048 is shown as SEQ ID No.48, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.288, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 528;
the nucleotide sequence of the forward primer 1 for detecting Ts049 is shown as SEQ ID No.49, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.289, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 529;
the nucleotide sequence of a forward primer 1 for detecting Ts050 is shown as SEQ ID No.50, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.290, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 530;
the nucleotide sequence of a forward primer 1 for detecting Ts051 is shown as SEQ ID No.51, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.291, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 531;
the nucleotide sequence of the forward primer 1 for detecting Ts052 is shown as SEQ ID No.52, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.292, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 532;
the nucleotide sequence of the forward primer 1 for detecting Ts053 is shown as SEQ ID No.53, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.293, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 533;
the nucleotide sequence of the forward primer 1 for detecting Ts054 is shown as SEQ ID No.54, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.294, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 534;
the nucleotide sequence of a forward primer 1 for detecting Ts055 is shown as SEQ ID No.55, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.295, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 535;
the nucleotide sequence of the forward primer 1 for detecting Ts056 is shown as SEQ ID No.56, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.296, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 536;
the nucleotide sequence of the forward primer 1 for detecting Ts057 is shown as SEQ ID No.57, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.297, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 537;
the nucleotide sequence of the forward primer 1 for detecting Ts058 is shown as SEQ ID No.58, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.298, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 538;
the nucleotide sequence of a forward primer 1 for detecting Ts059 is shown as SEQ ID No.59, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.299, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 539;
the nucleotide sequence of a forward primer 1 for detecting Ts060 is shown as SEQ ID No.60, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.300, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 540;
the nucleotide sequence of the forward primer 1 for detecting Ts061 is shown as SEQ ID No.61, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.301, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 541;
the nucleotide sequence of a forward primer 1 for detecting Ts062 is shown as SEQ ID No.62, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.302, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 542;
the nucleotide sequence of a forward primer 1 for detecting Ts063 is shown as SEQ ID No.63, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.303, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 543;
the nucleotide sequence of a forward primer 1 for detecting Ts064 is shown as SEQ ID No.64, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.304, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 544;
the nucleotide sequence of a forward primer 1 for detecting Ts065 is shown as SEQ ID No.65, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.305, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 545;
the nucleotide sequence of the forward primer 1 for detecting Ts066 is shown as SEQ ID No.66, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.306, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 546;
the nucleotide sequence of a forward primer 1 for detecting Ts067 is shown as SEQ ID No.67, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.307, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 547;
the nucleotide sequence of a forward primer 1 for detecting Ts068 is shown as SEQ ID No.68, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.308, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 548;
the nucleotide sequence of a forward primer 1 for detecting Ts069 is shown as SEQ ID No.69, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.309, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 549;
the nucleotide sequence of a forward primer 1 for detecting Ts070 is shown as SEQ ID No.70, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.310, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 550;
the nucleotide sequence of a forward primer 1 for detecting Ts071 is shown as SEQ ID No.71, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.311, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 551;
the nucleotide sequence of the forward primer 1 for detecting Ts072 is shown in SEQ ID No.72, the nucleotide sequence of the forward primer 2 is shown in SEQ ID No.312, and the nucleotide sequence of the reverse primer is shown in SEQ ID No. 552;
the nucleotide sequence of the forward primer 1 for detecting Ts073 is shown as SEQ ID No.73, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.313, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 553;
the nucleotide sequence of a forward primer 1 for detecting Ts074 is shown as SEQ ID No.74, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.314, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 554;
the nucleotide sequence of a forward primer 1 for detecting Ts075 is shown as SEQ ID No.75, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.315, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 555;
the nucleotide sequence of a forward primer 1 for detecting Ts076 is shown as SEQ ID No.76, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.316, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 556;
the nucleotide sequence of the forward primer 1 for detecting Ts077 is shown as SEQ ID No.77, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.317, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 557;
the nucleotide sequence of the forward primer 1 for detecting Ts078 is shown as SEQ ID No.78, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.318, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 558;
the nucleotide sequence of the forward primer 1 for detecting Ts079 is shown as SEQ ID No.79, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.319, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 559;
the nucleotide sequence of the forward primer 1 for detecting Ts080 is shown as SEQ ID No.80, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.320, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 560;
the nucleotide sequence of the forward primer 1 for detecting Ts081 is shown as SEQ ID No.81, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.321, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 561;
the nucleotide sequence of a forward primer 1 for detecting Ts082 is shown as SEQ ID No.82, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.322, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 562;
the nucleotide sequence of the forward primer 1 for detecting Ts083 is shown as SEQ ID No.83, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.323, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 563;
the nucleotide sequence of a forward primer 1 for detecting Ts084 is shown as SEQ ID No.84, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.324, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 564;
the nucleotide sequence of the forward primer 1 for detecting Ts085 is shown as SEQ ID No.85, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.325, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 565;
the nucleotide sequence of the forward primer 1 for detecting Ts086 is shown as SEQ ID No.86, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.326, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 566;
the nucleotide sequence of the forward primer 1 for detecting Ts087 is shown as SEQ ID No.87, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.327, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 567;
the nucleotide sequence of the forward primer 1 for detecting Ts088 is shown as SEQ ID No.88, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.328, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 568;
the nucleotide sequence of a forward primer 1 for detecting Ts089 is shown as SEQ ID No.89, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.329, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 569;
the nucleotide sequence of a forward primer 1 for detecting Ts090 is shown as SEQ ID No.90, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.330, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 570;
the nucleotide sequence of the forward primer 1 for detecting Ts091 is shown as SEQ ID No.91, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.331, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 571;
the nucleotide sequence of the forward primer 1 for detecting Ts092 is shown as SEQ ID No.92, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.332, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 572;
the nucleotide sequence of the forward primer 1 for detecting Ts093 is shown as SEQ ID No.93, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.333, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 573;
the nucleotide sequence of the forward primer 1 for detecting Ts094 is shown as SEQ ID No.94, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.334, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 574;
the nucleotide sequence of the forward primer 1 for detecting Ts095 is shown as SEQ ID No.95, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.335, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 575;
the nucleotide sequence of the forward primer 1 for detecting Ts096 is shown as SEQ ID No.96, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.336, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 576;
the nucleotide sequence of the forward primer 1 for detecting Ts097 is shown as SEQ ID No.97, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.337, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 577;
the nucleotide sequence of the forward primer 1 for detecting Ts098 is shown as SEQ ID No.98, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.338, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 578;
the nucleotide sequence of the forward primer 1 for detecting Ts099 is shown as SEQ ID No.99, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.339, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 579;
the nucleotide sequence of a forward primer 1 for detecting Ts100 is shown as SEQ ID No.100, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.340, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 580;
the nucleotide sequence of a forward primer 1 for detecting Ts101 is shown as SEQ ID No.101, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.341, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 581;
the nucleotide sequence of a forward primer 1 for detecting Ts102 is shown as SEQ ID No.102, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.342, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 582;
the nucleotide sequence of a forward primer 1 for detecting Ts103 is shown as SEQ ID No.103, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.343, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 583;
the nucleotide sequence of a forward primer 1 for detecting Ts104 is shown as SEQ ID No.104, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.344, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 584;
the nucleotide sequence of the forward primer 1 for detecting Ts105 is shown as SEQ ID No.105, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.345, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 585;
the nucleotide sequence of a forward primer 1 for detecting Ts106 is shown as SEQ ID No.106, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.346, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 586;
the nucleotide sequence of a forward primer 1 for detecting Ts107 is shown as SEQ ID No.107, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.347, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 587;
the nucleotide sequence of a forward primer 1 for detecting Ts108 is shown as SEQ ID No.108, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.348, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 588;
the nucleotide sequence of a forward primer 1 for detecting Ts109 is shown as SEQ ID No.109, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.349, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 589;
the nucleotide sequence of a forward primer 1 for detecting Ts110 is shown as SEQ ID No.110, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.350, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 590;
the nucleotide sequence of a forward primer 1 for detecting Ts111 is shown as SEQ ID No.111, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.351, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 591;
the nucleotide sequence of the forward primer 1 for detecting Ts112 is shown as SEQ ID No.112, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.352, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 592;
the nucleotide sequence of a forward primer 1 for detecting Ts113 is shown as SEQ ID No.113, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.353, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 593;
the nucleotide sequence of a forward primer 1 for detecting Ts114 is shown as SEQ ID No.114, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.354, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 594;
the nucleotide sequence of the forward primer 1 for detecting Ts115 is shown as SEQ ID No.115, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.355, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 595;
the nucleotide sequence of the forward primer 1 for detecting Ts116 is shown as SEQ ID No.116, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.356, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 596;
the nucleotide sequence of the forward primer 1 for detecting Ts117 is shown as SEQ ID No.117, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.357, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 597;
the nucleotide sequence of a forward primer 1 for detecting Ts118 is shown as SEQ ID No.118, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.358, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 598;
the nucleotide sequence of a forward primer 1 for detecting Ts119 is shown as SEQ ID No.119, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.359, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 599;
the nucleotide sequence of the forward primer 1 for detecting Ts120 is shown as SEQ ID No.120, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.360, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 600;
the nucleotide sequence of a forward primer 1 for detecting Ts121 is shown as SEQ ID No.121, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.361, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 601;
the nucleotide sequence of the forward primer 1 for detecting Ts122 is shown as SEQ ID No.122, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.362, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 602;
the nucleotide sequence of a forward primer 1 for detecting Ts123 is shown as SEQ ID No.123, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.363, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 603;
the nucleotide sequence of a forward primer 1 for detecting Ts124 is shown as SEQ ID No.124, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.364, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 604;
the nucleotide sequence of a forward primer 1 for detecting Ts125 is shown as SEQ ID No.125, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.365, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 605;
the nucleotide sequence of the forward primer 1 for detecting Ts126 is shown as SEQ ID No.126, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.366, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 606;
the nucleotide sequence of a forward primer 1 for detecting Ts127 is shown as SEQ ID No.127, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.367, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 607;
the nucleotide sequence of the forward primer 1 for detecting Ts128 is shown as SEQ ID No.128, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.368, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 608;
the nucleotide sequence of the forward primer 1 for detecting Ts129 is shown as SEQ ID No.129, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.369, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 609;
the nucleotide sequence of a forward primer 1 for detecting Ts130 is shown as SEQ ID No.130, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.370, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 610;
the nucleotide sequence of the forward primer 1 for detecting Ts131 is shown as SEQ ID No.131, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.371, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 611;
the nucleotide sequence of the forward primer 1 for detecting Ts132 is shown as SEQ ID No.132, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.372, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 612;
the nucleotide sequence of the forward primer 1 for detecting Ts133 is shown as SEQ ID No.133, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.373, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 613;
the nucleotide sequence of a forward primer 1 for detecting Ts134 is shown as SEQ ID No.134, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.374, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 614;
the nucleotide sequence of a forward primer 1 for detecting Ts135 is shown as SEQ ID No.135, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.375, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 615;
the nucleotide sequence of the forward primer 1 for detecting Ts136 is shown as SEQ ID No.136, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.376, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 616;
the nucleotide sequence of a forward primer 1 for detecting Ts137 is shown as SEQ ID No.137, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.377, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 617;
the nucleotide sequence of a forward primer 1 for detecting Ts138 is shown as SEQ ID No.138, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.378, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 618;
the nucleotide sequence of a forward primer 1 for detecting Ts139 is shown as SEQ ID No.139, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.379, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 619;
the nucleotide sequence of the forward primer 1 for detecting Ts140 is shown as SEQ ID No.140, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.380, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 620;
the nucleotide sequence of a forward primer 1 for detecting Ts141 is shown as SEQ ID No.141, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.381, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 621;
the nucleotide sequence of a forward primer 1 for detecting Ts142 is shown as SEQ ID No.142, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.382, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 622;
the nucleotide sequence of a forward primer 1 for detecting Ts143 is shown as SEQ ID No.143, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.383, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 623;
the nucleotide sequence of the forward primer 1 for detecting Ts144 is shown as SEQ ID No.144, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.384, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 624;
the nucleotide sequence of a forward primer 1 for detecting Ts145 is shown as SEQ ID No.145, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.385, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 625;
the nucleotide sequence of a forward primer 1 for detecting Ts146 is shown as SEQ ID No.146, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.386, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 626;
the nucleotide sequence of the forward primer 1 for detecting Ts147 is shown as SEQ ID No.147, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.387, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 627;
the nucleotide sequence of the forward primer 1 for detecting Ts148 is shown as SEQ ID No.148, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.388, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 628;
the nucleotide sequence of a forward primer 1 for detecting Ts149 is shown as SEQ ID No.149, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.389, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 629;
the nucleotide sequence of the forward primer 1 for detecting Ts150 is shown as SEQ ID No.150, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.390, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 630;
the nucleotide sequence of a forward primer 1 for detecting Ts151 is shown as SEQ ID No.151, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.391, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 631;
the nucleotide sequence of the forward primer 1 for detecting Ts152 is shown as SEQ ID No.152, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.392, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 632;
the nucleotide sequence of the forward primer 1 for detecting Ts153 is shown as SEQ ID No.153, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.393, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 633;
the nucleotide sequence of a forward primer 1 for detecting Ts154 is shown as SEQ ID No.154, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.394, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 634;
the nucleotide sequence of the forward primer 1 for detecting Ts155 is shown as SEQ ID No.155, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.395, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 635;
the nucleotide sequence of a forward primer 1 for detecting Ts156 is shown as SEQ ID No.156, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.396, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 636;
the nucleotide sequence of a forward primer 1 for detecting Ts157 is shown as SEQ ID No.157, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.397, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 637;
the nucleotide sequence of a forward primer 1 for detecting Ts158 is shown as SEQ ID No.158, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.398, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 638;
the nucleotide sequence of a forward primer 1 for detecting Ts159 is shown as SEQ ID No.159, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.399, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 639;
the nucleotide sequence of the forward primer 1 for detecting Ts160 is shown as SEQ ID No.160, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.400, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 640;
the nucleotide sequence of a forward primer 1 for detecting Ts161 is shown as SEQ ID No.161, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.401, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 641;
the nucleotide sequence of the forward primer 1 for detecting Ts162 is shown as SEQ ID No.162, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.402, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 642;
the nucleotide sequence of a forward primer 1 for detecting Ts163 is shown as SEQ ID No.163, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.403, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 643;
the nucleotide sequence of a forward primer 1 for detecting Ts164 is shown as SEQ ID No.164, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.404, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 644;
the nucleotide sequence of the forward primer 1 for detecting Ts165 is shown as SEQ ID No.165, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.405, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 645;
the nucleotide sequence of a forward primer 1 for detecting Ts166 is shown as SEQ ID No.166, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.406, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 646;
the nucleotide sequence of the forward primer 1 for detecting Ts167 is shown as SEQ ID No.167, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.407, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 647;
the nucleotide sequence of a forward primer 1 for detecting Ts168 is shown as SEQ ID No.168, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.408, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 648;
the nucleotide sequence of the forward primer 1 for detecting Ts169 is shown as SEQ ID No.169, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.409, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 649;
the nucleotide sequence of a forward primer 1 for detecting Ts170 is shown as SEQ ID No.170, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.410, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 650;
the nucleotide sequence of a forward primer 1 for detecting Ts171 is shown as SEQ ID No.171, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.411, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 651;
the nucleotide sequence of the forward primer 1 for detecting Ts172 is shown as SEQ ID No.172, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.412, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 652;
the nucleotide sequence of a forward primer 1 for detecting Ts173 is shown as SEQ ID No.173, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.413, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 653;
the nucleotide sequence of the forward primer 1 for detecting Ts174 is shown as SEQ ID No.174, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.414, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 654;
the nucleotide sequence of a forward primer 1 for detecting Ts175 is shown as SEQ ID No.175, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.415, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 655;
the nucleotide sequence of the forward primer 1 for detecting Ts176 is shown as SEQ ID No.176, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.416, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 656;
the nucleotide sequence of the forward primer 1 for detecting Ts177 is shown as SEQ ID No.177, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.417, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 657;
the nucleotide sequence of a forward primer 1 for detecting Ts178 is shown as SEQ ID No.178, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.418, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 658;
the nucleotide sequence of the forward primer 1 for detecting Ts179 is shown as SEQ ID No.179, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.419, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 659;
the nucleotide sequence of a forward primer 1 for detecting Ts180 is shown as SEQ ID No.180, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.420, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 660;
the nucleotide sequence of a forward primer 1 for detecting Ts181 is shown as SEQ ID No.181, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.421, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 661;
the nucleotide sequence of a forward primer 1 for detecting Ts182 is shown as SEQ ID No.182, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.422, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 662;
the nucleotide sequence of a forward primer 1 for detecting Ts183 is shown as SEQ ID No.183, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.423, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 663;
the nucleotide sequence of the forward primer 1 for detecting Ts184 is shown as SEQ ID No.184, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.424, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 664;
the nucleotide sequence of a forward primer 1 for detecting Ts185 is shown as SEQ ID No.185, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.425, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 665;
the nucleotide sequence of the forward primer 1 for detecting Ts186 is shown as SEQ ID No.186, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.426, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 666;
the nucleotide sequence of a forward primer 1 for detecting Ts187 is shown as SEQ ID No.187, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.427, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 667;
the nucleotide sequence of a forward primer 1 for detecting Ts188 is shown as SEQ ID No.188, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.428, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 668;
the nucleotide sequence of a forward primer 1 for detecting Ts189 is shown as SEQ ID No.189, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.429, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 669;
the nucleotide sequence of a forward primer 1 for detecting Ts190 is shown as SEQ ID No.190, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.430, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 670;
the nucleotide sequence of a forward primer 1 for detecting Ts191 is shown as SEQ ID No.191, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.431, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 671;
the nucleotide sequence of a forward primer 1 for detecting Ts192 is shown as SEQ ID No.192, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.432, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 672;
the nucleotide sequence of the forward primer 1 for detecting Ts193 is shown as SEQ ID No.193, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.433, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 673;
the nucleotide sequence of a forward primer 1 for detecting Ts194 is shown as SEQ ID No.194, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.434, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 674;
the nucleotide sequence of a forward primer 1 for detecting Ts195 is shown as SEQ ID No.195, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.435, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 675;
the nucleotide sequence of a forward primer 1 for detecting Ts196 is shown as SEQ ID No.196, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.436, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 676;
the nucleotide sequence of a forward primer 1 for detecting Ts197 is shown as SEQ ID No.197, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.437, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 677;
the nucleotide sequence of the forward primer 1 for detecting Ts198 is shown as SEQ ID No.198, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.438, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 678;
the nucleotide sequence of a forward primer 1 for detecting Ts199 is shown as SEQ ID No.199, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.439, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 679;
the nucleotide sequence of the forward primer 1 for detecting Ts200 is shown as SEQ ID No.200, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.440, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 680;
the nucleotide sequence of a forward primer 1 for detecting Ts201 is shown as SEQ ID No.201, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.441, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 681;
the nucleotide sequence of a forward primer 1 for detecting Ts202 is shown as SEQ ID No.202, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.442, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 682;
the nucleotide sequence of a forward primer 1 for detecting Ts203 is shown as SEQ ID No.203, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.443, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 683;
the nucleotide sequence of a forward primer 1 for detecting Ts204 is shown as SEQ ID No.204, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.444, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 684;
the nucleotide sequence of a forward primer 1 for detecting Ts205 is shown as SEQ ID No.205, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.445, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 685;
the nucleotide sequence of a forward primer 1 for detecting Ts206 is shown as SEQ ID No.206, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.446, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 686;
the nucleotide sequence of a forward primer 1 for detecting Ts207 is shown as SEQ ID No.207, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.447, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 687;
the nucleotide sequence of the forward primer 1 for detecting Ts208 is shown as SEQ ID No.208, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.448, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 688;
the nucleotide sequence of a forward primer 1 for detecting Ts209 is shown as SEQ ID No.209, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.449, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 689;
the nucleotide sequence of the forward primer 1 for detecting Ts210 is shown as SEQ ID No.210, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.450, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 690;
the nucleotide sequence of the forward primer 1 for detecting Ts211 is shown as SEQ ID No.211, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.451, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 691;
the nucleotide sequence of the forward primer 1 for detecting Ts212 is shown as SEQ ID No.212, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.452, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 692;
the nucleotide sequence of a forward primer 1 for detecting Ts213 is shown as SEQ ID No.213, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.453, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 693;
the nucleotide sequence of the forward primer 1 for detecting Ts214 is shown as SEQ ID No.214, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.454, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 694;
the nucleotide sequence of a forward primer 1 for detecting Ts215 is shown as SEQ ID No.215, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.455, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 695;
the nucleotide sequence of a forward primer 1 for detecting Ts216 is shown as SEQ ID No.216, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.456, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 696;
the nucleotide sequence of a forward primer 1 for detecting Ts217 is shown as SEQ ID No.217, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.457, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 697;
the nucleotide sequence of a forward primer 1 for detecting Ts218 is shown as SEQ ID No.218, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.458, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 698;
the nucleotide sequence of a forward primer 1 for detecting Ts219 is shown as SEQ ID No.219, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.459, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 699;
the nucleotide sequence of a forward primer 1 for detecting Ts220 is shown as SEQ ID No.220, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.460, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 700;
the nucleotide sequence of the forward primer 1 for detecting Ts221 is shown as SEQ ID No.221, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.461, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 701;
the nucleotide sequence of the forward primer 1 for detecting Ts222 is shown as SEQ ID No.222, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.462, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 702;
the nucleotide sequence of a forward primer 1 for detecting Ts223 is shown as SEQ ID No.223, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.463, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 703;
the nucleotide sequence of a forward primer 1 for detecting Ts224 is shown as SEQ ID No.224, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.464, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 704;
the nucleotide sequence of a forward primer 1 for detecting Ts225 is shown as SEQ ID No.225, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.465, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 705;
the nucleotide sequence of a forward primer 1 for detecting Ts226 is shown as SEQ ID No.226, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.466, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 706;
the nucleotide sequence of the forward primer 1 for detecting Ts227 is shown as SEQ ID No.227, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.467, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 707;
the nucleotide sequence of the forward primer 1 for detecting Ts228 is shown as SEQ ID No.228, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.468, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 708;
the nucleotide sequence of a forward primer 1 for detecting Ts229 is shown as SEQ ID No.229, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.469, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 709;
the nucleotide sequence of the forward primer 1 for detecting Ts230 is shown as SEQ ID No.230, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.470, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 710;
the nucleotide sequence of a forward primer 1 for detecting Ts231 is shown as SEQ ID No.231, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.471, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 711;
the nucleotide sequence of a forward primer 1 for detecting Ts232 is shown as SEQ ID No.232, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.472, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 712;
the nucleotide sequence of the forward primer 1 for detecting Ts233 is shown as SEQ ID No.233, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.473, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 713;
the nucleotide sequence of a forward primer 1 for detecting Ts234 is shown as SEQ ID No.234, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.474, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 714;
the nucleotide sequence of a forward primer 1 for detecting Ts235 is shown as SEQ ID No.235, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.475, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 715;
the nucleotide sequence of the forward primer 1 for detecting Ts236 is shown as SEQ ID No.236, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.476, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 716;
the nucleotide sequence of a forward primer 1 for detecting Ts237 is shown in SEQ ID No.237, the nucleotide sequence of a forward primer 2 is shown in SEQ ID No.477, and the nucleotide sequence of a reverse primer is shown in SEQ ID No. 717;
the nucleotide sequence of a forward primer 1 for detecting Ts238 is shown as SEQ ID No.238, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.478, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 718;
the nucleotide sequence of a forward primer 1 for detecting Ts239 is shown as SEQ ID No.239, the nucleotide sequence of a forward primer 2 is shown as SEQ ID No.479, and the nucleotide sequence of a reverse primer is shown as SEQ ID No. 719;
the nucleotide sequence of the forward primer 1 for detecting Ts240 is shown as SEQ ID No.240, the nucleotide sequence of the forward primer 2 is shown as SEQ ID No.480, and the nucleotide sequence of the reverse primer is shown as SEQ ID No. 720.
3. A set of KASP primers according to claim 2, wherein: the 5' end of the forward primer 1 is connected with a FAM fluorescent label sequence;
the 5' end of the forward primer 2 is connected with a HEX fluorescent tag sequence.
4. The set of KASP primers of claim 3, wherein: the FAM fluorescent label sequence is shown as SEQ ID No. 721;
the HEX fluorescent tag sequence is shown as SEQ ID No. 722.
5. A kit comprising a set of KASP primers according to any one of claims 2 to 4.
6. Use of a set of KASP primers according to any of claims 2 to 4 or a kit according to claim 5 in any of:
(1) Preparing a kit for detecting the genotype of the SNP locus of the tobacco;
(2) Genotyping the tobacco germplasm resources or genetic groups;
(3) Constructing a DNA fingerprint of tobacco germplasm resources or varieties;
(4) Detecting the purity or the authenticity of the tobacco variety;
the type of tobacco is selected from flue-cured tobacco, sun-cured tobacco, burley tobacco, cigar tobacco, aromatic tobacco and medicinal tobacco.
7. A method of genotyping a tobacco germplasm resource or genetic population comprising the steps of:
(a1) Respectively extracting genome DNA of all tobacco samples in the tested tobacco germplasm resources or genetic groups;
(a2) Preparing a PCR reaction system for the DNA of the tobacco sample to be tested obtained in the step (a 1) according to 240 SNP sites in claim 1, and respectively carrying out PCR amplification by using corresponding primers in the KASP primer set according to claim 3 or 4;
(a3) Detecting FAM and HEX fluorescence values of the PCR amplification product of the DNA of the tobacco sample to be tested at the 240 SNP sites in claim 1 by using a KASP fluorescence detector, and calculating to obtain the genotyping result of the DNA of the tobacco sample to be tested at the 240 SNP sites;
the type of tobacco is selected from flue-cured tobacco, sun-cured tobacco, burley tobacco, cigar tobacco, aromatic tobacco and medicinal tobacco.
8. A method for constructing a DNA fingerprint of a tobacco germplasm resource or variety comprises the following steps:
(b1) Extracting genome DNA of all tobacco germplasm resources or varieties for constructing a DNA fingerprint, detecting the genotypes of the 240 SNP sites in claim 1, and combining and arranging the genotypes of the 240 SNP sites of each tobacco germplasm resource or variety according to the site numbering sequence to form a specific sequence, wherein the specific sequence is the DNA fingerprint of the tobacco germplasm resources or varieties;
(b2) Selecting total 48 loci of Ts003, ts007, ts013, ts019, ts028, ts029, ts033, ts039, ts041, ts048, ts055, ts060, ts062, ts070, ts074, ts079, ts082, ts089, ts093, ts098, ts106, ts108, ts113, ts120, ts122, ts129, ts132, ts138, ts141, ts148, ts152, ts159, ts164, ts169, ts172, ts183, ts186, ts192, ts194, ts201, ts207, SNP 212, ts218, ts222, ts230, ts232 and Ts236 from the chromosome 1 to the chromosome 24 in sequence according to the principle of 2 SNP loci of each chromosome from the 240 SNP loci described in claim 1 as the core characteristic loci of the tobacco germplasm or DNA fingerprint map;
(b3) And (c) aiming at the 48 core characteristic SNP loci in the step (b 2), converting the genotype combination into a computer character string label according to the following steps:
firstly, dividing 48 core characteristic SNP loci into 24 pairs of SNP locus combinations according to a rule of combining SNP loci on the same chromosome, wherein the genotyping result of each SNP locus consists of XX, YY, XY or-wherein X and Y are single nucleotides of the locus and are A, T, G or C, and-represents deletion or invalid data, two SNP loci in each pair of SNP locus combinations are arranged in sequence from small to large according to the physical position on the chromosome, and each pair of SNP locus genotypes are combined with 16 homozygous genotype combinations of AAAA, AATT, AAGG, AACC, TTTT, TTGG, TTCC, GGAA, GGTT, GGGGCC, CCAA, CCCCTT, CCGG and CCCC and genotype combinations comprising XY or-types;
then, converting each pair of SNP locus homozygous genotype combinations according to the character '0' corresponding to AAAA, the character '1' corresponding to AATT, ' the character '2' corresponding to AAGG, the character '3' corresponding to AACC, the character '4' corresponding to TTAA, the character '5' corresponding to TTTT, the character '6' corresponding to TTGG, the character '7' corresponding to TTCC, the character '8' corresponding to GGAA, the character '9' corresponding to GGTT, the character ' a ' corresponding to GGCC, the character ' b ' corresponding to CCAA, the character'd ' corresponding to CCTT, the character ' e ' corresponding to CCGG and the character ' f ' corresponding to CCCC, defining the genotype combination containing XY or-type as ' nonsense ', uniformly converting the genotype combination into the character ' n ', and sequentially converting the 24 pairs of SNP locus combinations according to chromosome numbers;
according to the rules, the genotype combination of the 48 core characteristic SNP loci in the DNA fingerprint of the tobacco germplasm resource or variety is converted into a character string consisting of 24 characters, and the character string is the core code of the DNA fingerprint of the tobacco germplasm resource or variety;
(b4) On the basis of the core code of the DNA fingerprint of the tobacco germplasm resource or variety, the genotype data of the 240 SNP sites in the claim 1 is combined, and additional code and annotation code segments are added, wherein the format is as follows: a core code-additional code-annotation code, wherein the core code is a character string composed of 24 characters defined in (b 3), the additional code is 1 of the characters 0-9, a-f and n, and the annotation code is a character string composed of the characters 0-9 and having a length of 6;
the additional code is the coding of the genotype of any two SNP loci which can distinguish the two SNP loci of the tested tobacco germplasm resources or varieties except the 48 core characteristic SNP loci in the 240 SNP loci in the claim 1, and the coding rule is the same as that in the step (b 3);
the annotation code is the sequence numbers of two SNP sites corresponding to the additional code;
(b5) Converting the DNA fingerprint core code or core code-additional code-annotation code characters of the tobacco germplasm resources or varieties constructed in the step (b 4) into a two-dimensional code or bar code which can be identified by an equipment terminal so as to intelligently identify and manage the tobacco germplasm resources or varieties;
the type of tobacco is selected from flue-cured tobacco, sun-cured tobacco, burley tobacco, cigar tobacco, aromatic tobacco and medicinal tobacco.
9. A method for detecting the purity or the authenticity of a tobacco variety comprises the following steps:
(c1) Preparing more than 50 seeds of a tobacco variety to be detected and a standard control tobacco variety respectively, accelerating germination and sowing, taking an equivalent tissue of a single seedling when the seedling grows out of 4 true leaves, dividing the seedling into two groups according to the tobacco variety to be detected and the standard control tobacco variety, mixing and preparing samples, and extracting genome DNA;
(c2) Detecting the genotypes of the 240 SNP sites in the claim 1, thereby obtaining the genotype data of the tobacco variety to be detected and the standard control tobacco variety at the 240 SNP sites; when the genotype of the standard control tobacco variety sample at a certain SNP locus is XX and YY, the locus is an effective locus, and if the genotype of the locus is XY or-type, the locus is an ineffective locus; wherein X, Y are A, T, G or C, -represent missing or invalid data;
(c3) Comparing the genotype difference between the tobacco variety to be tested and the standard control tobacco variety with respect to the effective locus of the standard control tobacco variety sample in the 240 SNP loci in claim 1, wherein the two results are as follows: the genotypes are the same or different;
(c4) Variety purity: (P) The value of (c) is expressed in%, calculated according to equation (1):
Figure 2
in the formula:
P-variety purity;
s1, the number of effective loci of the standard control tobacco variety;
s2, in the effective sites of the standard comparison tobacco variety, the number of the sites with different genotypes of the tobacco variety to be detected and the standard comparison tobacco variety is counted;
(c5) When the purity of the tobacco variety to be detected is less than or equal to 95%, judging that the tobacco variety to be detected and the standard contrast variety are different varieties, when the purity of the tobacco variety to be detected is more than 95% and less than 100%, judging that the tobacco variety to be detected is an approximate variety or a suspected same variety of the standard contrast variety, and when the purity of the tobacco variety to be detected is 100%, judging that the tobacco variety to be detected and the standard contrast variety are the same variety;
the tobacco is selected from flue-cured tobacco, sun-cured tobacco, burley tobacco, cigar, aromatic tobacco, and medicinal tobacco.
10. The method according to any one of claims 7-9, wherein: in the method, the complete set of KASP primers of any one of claims 2-4 or the kit of claim 5 is used for completing the genotype detection of the 240 SNP sites.
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