CN112080570A - KASP labeled primer combination for identifying hybrid stichopus japonicus in Zhongrussia and application thereof - Google Patents
KASP labeled primer combination for identifying hybrid stichopus japonicus in Zhongrussia and application thereof Download PDFInfo
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Abstract
The invention relates to a KASP labeled primer combination for identifying hybrid stichopus japonicus and an application thereof, belonging to the field of molecular biology, wherein the primer combination is shown as SEQ ID NO. 1-30. The invention also provides a method for identifying the population and the individual of the filial generation stichopus japonicus in Russian or the filial generation stichopus japonicus in different batches by using the primer combination, which comprises the following specific steps: extracting DNA of a stichopus japonicus sample by adopting an SDS method, carrying out SNP genotyping on the sample to be detected by using a KASP marker, wherein the judgment standard is as follows: the population detection rate reaches 70 percent, and is a Chinese and Russian hybrid stichopus japonicus population- 'Water institute No. 1' stichopus japonicus population, and the population detection rate is the proportion of the number of individuals with heterozygosis detected by at least 1 primer in the population to the total detected individuals; wherein, the individual sample covered by at least 3 markers is filial generation stichopus japonicus or the cross stichopus japonicus in different batches, namely the individual 'Water institute No. 1'. The labeled primer can successfully identify the hybrid stichopus japonicus, and the identification rate is 100%.
Description
Technical Field
The invention belongs to the field of molecular biology, and particularly relates to a KASP labeled primer combination for identifying hybrid stichopus japonicus and application thereof.
Background
Paternity testing, also called paternity testing, is a technology for detecting and analyzing the genetic relationship between parents and filial generations by using technical means such as molecular biology, genetics and the like, and judging whether genetic relationship exists or not from the aspects of molecular heredity, appearance structure, physiological function and the like of the filial generations and the parents. Paternity test is also significant in aquatic animal genetic breeding, and can be used for judging hybrid or invisible species; through paternity test, the genetic relationship of the offspring obtained by group breeding can be determined, and the reduction of genetic diversity caused by inbreeding can be effectively avoided; meanwhile, the molecular marker information in the paternity test process can also be used for analyzing the genetic distance and the genetic variation level between parents and filial generations and guiding population breeding. At present, molecular markers become the first choice for paternity test analysis due to the characteristics of sensitivity, stability, abundant quantity, accuracy, reliability and the like. Compared with traditional molecular markers (RFLP, RAPD and SSR), the Single nucleotide polymorphism marker (SNP) has the characteristics of high polymorphism, wide distribution, strong genetic stability and the like, can realize high-throughput and automatic detection, has the advantages of low cost and high efficiency, and plays an important role in paternity test and genetic breeding.
In the aquatic economic animal breeding technology, crossbreeding can concentrate two or more excellent characters in the same species into a new variety, and obtain a new variety (generating heterosis) stronger or better than a parent variety. The crossbreeding technology is simple to operate, is suitable for aquatic animals to lay eggs and discharge sperm in large quantity, can directly utilize the production mode of the first filial generation heterosis, and is widely applied to the fine breed cultivation of the aquatic animals. The hybrid stichopus japonicus 'Water institute No. 1' cultivated by Dalian sea university is cultivated by taking Russian sea cucumber river-like (Helady Stoke) stichopus japonicus and Liaoning (Zhuang river) stichopus japonicus as male and female parents through 'selection from top to bottom'. The hybrid stichopus japonicus in China and Russia has the characteristics of large individual, much thorns and strong tolerance of the stichopus japonicus in Russia and has the characteristics of: the number of wart feet (meat spurs) on the body surface of the hybrid stichopus japonicus is large, and the hybrid stichopus japonicus is arranged in 6 rows in order; the body wall thickness is increased by more than 10 percent compared with the control group of the Chinese stichopus japonicus population by the body wall weight/living body weight (meat yield and skin yield); the culture survival rate is high, the growth speed is improved by more than 30 percent, and the pond culture yield is high. At present, the hybrid stichopus japonicus and stichopus japonicus in the areas of Liaoning, Shandong, Hebei, and the like are popularized and demonstrated, the economic benefit of cultivation is remarkable, and the cost of the stichopus japonicus and stichopus japonicus seedlings is higher than that of the common stichopus japonicus and stichopus japonicus seedlings. Therefore, the market is disturbed by the appearance of counterfeit hybrid stichopus japonicus seedlings in the market, and farmers suffer huge losses.
Disclosure of Invention
The invention aims to solve the technical problem of providing a KASP marker primer combination for identifying the Chinese and Russian hybrid stichopus japonicus 'Water park No. 1' and an application method thereof, wherein the marker can successfully identify the Chinese and Russian hybrid stichopus japonicus 'Water park No. 1'.
The invention is realized by the following technical scheme:
a KASP labeled primer combination for identifying hybrid stichopus japonicus in Zhongrussia is shown in Table 1;
TABLE 110 KASP-tagged primer combinations for the detection of Russian-hybridized Stichopus japonicus
The invention also provides an application of the KASP labeled primer combination, and the application is specifically to the application of the primer combination to identify the filial generation stichopus japonicus or the population and individuals of the Russian hybrid stichopus japonicus in different batches, and the specific method is as follows: extracting DNA of a stichopus japonicus sample by adopting an SDS method, carrying out SNP genotyping on the sample to be detected by using a KASP marker primer, wherein the judgment standard is as follows: the population detection rate reaches 70 percent, and is a Chinese and Russian hybrid stichopus japonicus population, and the population detection rate is the proportion of the number of individuals with at least 1 primer detected as a heterozygous type in the population to the total detected individuals; wherein the individual sample covered by at least 3 markers is filial stichopus japonicus or cross stichopus japonicus in different batches; the hybrid Chinese and Russian stichopus japonicus is a Water institute No.1 individual.
Detailed Description
The technical solution of the present invention is further explained by the following examples, but the scope of the present invention is not limited in any way by the examples.
Example 1
The research uses the whole genome sequence of the stichopus japonicus as a reference, carries out individual re-sequencing on the female parent of the stichopus japonicus for Liaoning village river and the male parent of the Russian stichopus japonicus for river sprain respectively, and obtains SNP marker loci of the stichopus japonicus population and the Russian stichopus japonicus population by comparing the SNP marker loci with the reference genome; and finally obtaining the KASP marker which can be used for identifying the filial generation stichopus japonicus or the filial generation stichopus japonicus in different batches, namely the individual 'Water institute No. 1'.
The specific method comprises the following steps:
1. mark acquisition
1.1DNA sample extraction
Selecting 5 representative and good-state 5 Chinese Liaoning village river stichopus japonicus (female parent), 5 Russian sea cucumber (river stichopus japonicus) (male parent) which are individuals obtained by respectively expanding propagation of the first generation parent of the stichopus japonicus of Water institute No. 1) and 5 filial generations of the stichopus japonicus, and extracting the genomic DNA of the stichopus japonicus by adopting an SDS method, wherein the DNA concentration is more than 100 ng/ul.
1.2 individual genome resequencing
The procedures of sample quality detection, library construction, library quality detection, library sequencing and the like are all executed according to the protocol standard (second generation high throughput sequencing provided by Illumina). And (3) carrying out DNA fragmentation on a qualified sample by adopting an ultrasonic method, after purification, carrying out end repair, adding A at the 3' end, connecting a sequencing joint, then selecting fragments with different sizes by utilizing agarose gel electrophoresis, and carrying out PCR amplification to construct a sequencing library. Performing genome re-sequencing on the library qualified by quality inspection on an Illumina platform, counting the number of sequences of original double-end sequence reads obtained by sequencing in a unit of every 4 lines, and performing quality evaluation, wherein the evaluation principle is as follows:
(1) filtering out SNPs within 5bp near the INDEL and sites of adjacent INDEL within 10 bp;
(2) filtering out sites with a probability of having variant variation below 30 (i.e.quality > Q30%);
(3) filtering out sites with a low ratio of Quality of variation (Quality) to Depth of coverage (Depth) (i.e., QD > 2.0);
(4) filtering out sites with lower alignment quality values (i.e., MQ > 40);
(5) sites were filtered out where there was significant plus-minus strand specificity for reads containing only the variation and reads containing only the base of the reference sequence when sequenced or aligned (i.e., FS < 60.0).
The filtered Clean Reads were aligned to the reference genomic sequence. According to the positioning result of Clean Reads in a reference genome, redundant Reads are filtered by Picard, and then variation detection of SNP and InDel is carried out by using a HaplotypeCaller (local haplotype assembly) algorithm of GATK, so that 3 final variation sites of stichopus japonicus are obtained.
1.3 typing and screening of specific SNP sites
And summarizing all the different variation sites between the samples according to the comparison result of the samples and the reference genome, and screening 60 high-quality SNP sites for subsequent KASP verification. The screening principle is as follows: (1) the locus is homozygosis and has different genotypes in both parents, and is heterozygosis in filial generation (for example, the genotypes of the parents are AA and TT, and the genotype of the filial generation is AT); (2) adjacent variant sites can not be too close, and the distance is more than 50 bp; (3) the upstream and downstream sequences of the site have uniqueness in the genome, and less repeated sequences exist near the site. Extracting 200bp sequences upstream and downstream of the high-quality SNP locus to be developed into a marker, and obtaining the information of the screened SNP locus shown in the attached table 1.
1.4KASP tag design
According to the SNP sites, 60 pairs of PCR 3' end specific amplification primers are designed, the design software is Primer 5, and each sequence needs 3 primers, namely two specific upstream primers and one universal downstream Primer. The principle of primer design is as follows: (1) the primers do not have hairpin structures, and primer dimers cannot be formed between the upstream primers and the downstream primers; (2) the GC content of the primer is 40-60%, and the Tm value is 55-65 ℃.
Verification of KASP tags
2.1 validation of population DNA extraction and marker detection
Randomly selecting 30 Russian hybrid Stichopus japonicus in different batches (15 of them are labeled source population hybrid progeny individuals, and 15 of them are Russian hybrid Stichopus japonicus in different batches, namely "Water institute No. 1" Stichopus japonicus), extracting DNA by SDS method, and performing DNA extraction in Douglas Array
The method comprises the steps of carrying out label detection on Platform, carrying out fluorescent quantitative PCR reaction in a SOELLEX high-flux PCR water bath environment, and selecting TB Green as a reagentTM Premix Ex TaqTMII, preparing 20 mul mixed system to carry out specific amplification by a two-step reaction procedure of 'denaturation-annealing extension'.
2.2KASP marker typing
Signals of two fluorophores, namely FAM and VIC, are read by an ARAYA fluorescence reader, and the results are led into a database to carry out SNP typing on the sample on the Douglas Scientific Dashboard according to the principles of definite typing, NTC (no sample negative control) and no specific amplification. And analyzing the typing result by utilizing SNP viewer genotype reading software, and reading the heterozygous genotype by the KASP marker in the sample to obtain the successful typing. In the typing scheme, X: X represents homozygous type including five types A: A, T: T, C: C, G: G, - (deletion), and X: Y represents heterozygous type including three types A: G, C: T, A: - (partial deletion)? Representing no or weak signal, and uncalloble representing signal but no explicit typing. Finally, 25 KASP markers for successful typing were obtained.
Screening 10 markers from 60 KASP markers under the condition that the detection rate of the population (namely the proportion of detecting heterozygote by at least 1 marker in each individual of the population) reaches 100%; and each sample is covered by at least 5 tags, the sequences of 10 KASP tags are shown in Table 2.
3. Identification of hybrid Stichopus japonicus
The verification and identification are carried out on 10 filial generation stichopus japonicus populations, 30 Russian hybrid stichopus japonicus cultivated in different batches and 20 common Chinese stichopus japonicus, and the results show that:
(1) detecting the filial generation stichopus japonicus population, wherein the detection rate of the labeled population is 100%; a minimum of 7 markers covered per sample;
(2) the Russian hybrid stichopus japonicus ('Water institute No. 1') in different batches is detected, the detection rate of a labeled population is 100%, and each sample is covered by at least 4 labels;
(3) the method is used for detecting the common Chinese stichopus japonicus, the group detection rate is 0%, and no sample is covered by a label.
The judgment standard is as follows: carrying out SNP genotyping on a sample to be detected by using KASP markers, wherein the population detectable rate reaches 70%, and is a Chinese and Russian hybrid stichopus japonicus population- 'Water institute No. 1' stichopus japonicus population, and the population detectable rate is the proportion of the number of individuals detected by at least 1 marker in the population to the total detected individuals; wherein, the individual sample covered by at least 3 markers is filial generation stichopus japonicus or the cross stichopus japonicus in different batches, namely the individual 'Water institute No. 1'.
TABLE 210 high quality SNP site sequence information
Remarking: SNP sites are indicated by [ ].
Sequence listing
<110> university of Dalian ocean
<120> marking primer combination for identifying hybrid stichopus japonicus in Chinese and Russian provinces and application thereof
<160> 40
<170> SIPOSequenceListing 1.0
<210> 1
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
gctttggtgg agttcttt 18
<210> 2
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
ggtcggtatt atttgtgaa 19
<210> 3
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
tgactgaccg actgagaag 19
<210> 4
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 4
tggtgatggg tactgatg 18
<210> 5
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 5
gatgggtact gatgttgaa 19
<210> 6
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 6
caacaaagat agcaacctc 19
<210> 7
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 7
gagaatggta tcccagcaa 19
<210> 8
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 8
aaccaagtac aatgtaggac c 21
<210> 9
<211> 17
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 9
aaagcatccc gtgtctg 17
<210> 10
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 10
gagttacccc tctttcacc 19
<210> 11
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 11
caccatctgg caataaat 18
<210> 12
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 12
aacggtgtta ttgtcattct t 21
<210> 13
<211> 17
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 13
cttccccacc accactt 17
<210> 14
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 14
aagtaaaccc gtccatcc 18
<210> 15
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 15
ccacctaaca accaaacaa 19
<210> 16
<211> 17
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 16
taaagggcaa ccaaaag 17
<210> 17
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 17
aagtccctta gccagttc 18
<210> 18
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 18
aagacgaata ccaaagaaa 19
<210> 19
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 19
agcctttcta ttcccagta 19
<210> 20
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 20
tcagaaaatt cccgctat 18
<210> 21
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 21
atgcttgtat gtgcttctg 19
<210> 22
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 22
gaaaataccc tcccaaaa 18
<210> 23
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 23
acaggatctg gaaaatacc 19
<210> 24
<211> 16
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 24
ttccgaccaa accaca 16
<210> 25
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 25
ctggttacct ccataaagtc 20
<210> 26
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 26
ccttccagaa catcctca 18
<210> 27
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 27
tccaagttac ctttcagttt 20
<210> 28
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 28
tataaagccc cgtagtagc 19
<210> 29
<211> 18
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 29
tcggatcaac ttgtttca 18
<210> 30
<211> 21
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 30
agtagggaca gaatacagac a 21
<210> 31
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 31
aaacccaaga tgtttacatt gctgctctgt taagttttgt gtattttggt cattcatgat 60
tggtcggtat tatttgtgaa cctactagtg attcagtctc wataactcat cgaccgaggt 120
ttacatttct tctagaattc agaagctcag catcatgttg aactttacct ttcaagcact 180
gaagtcattt aaagatataa c 201
<210> 32
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 32
tttagcttct ggtgaacaac aagattatga gtttggactc cctgtaaaag aagtttacaa 60
gaagatctgg tgtccagata atgaggtgag acctgtgaag wtaacaatgt tgctctttcc 120
aattgtcttg gttttacttg aagtacaact gaagcacacc ctagtactgc tttaaaagta 180
gatatatgta tatatgcata t 201
<210> 33
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 33
tattagagaa tggtatccca gcaaaggggg atgaactgta ccaagcagct atgatagcag 60
tcagacgtaa ccaagtacaa tgtaggacca acaggtgggt sgataaactc aaaactcaaa 120
gatgacagac acgggatgct tttatcttga tccctcttca caacagtcta aatataaaat 180
tttgttactt taatggtgtc g 201
<210> 34
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 34
aaaaattaag aagctgtacg ttaattacac caattgtata gctgtctgta agtctacaca 60
ataaaagtaa aatcaaatca aatctaaagt tcttagaaga wtgaacaact gggttagtat 120
tgtagaaaaa atattgaatg aatgaattaa ttaattagta attaattaat aatagtgcga 180
tcagtgccag tatacactta a 201
<210> 35
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 35
tcacggcaca cctacagcag agtaaactag agctgtgttt ttagaacttt ctttcatatt 60
aacaatgtta atagttacag gtcaagtaat ggtgcaatgt wgaccataca tggtgcactg 120
taagaccaca catggtgcaa tcagagacca tatatggtgt actgtgcaac catgcatggt 180
gcactgtttg actacagact t 201
<210> 36
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 36
ttattatgca gagtacatca ttgcattctg tataaagggc aaccaaaagt cccttagcca 60
gttcatttca gatttatcac gtttgatagg attgcatttt stcacttgag attgtcttgg 120
ataaataaat tacaagtttc tttggtattc gtcttcaaag atactgtcta ttcttagttt 180
actgaattga aaagatttca c 201
<210> 37
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 37
tggttcaaat tgtttacatg tcatcaagca gccaacaatg ttactgacac aataaaacta 60
ttcagtagtt ttctgtgaat aagcaaacac aaaacacatg wtacaattgc tcctttatgg 120
aaactaatgt acaatccaga agcacataca agcatcagtt tatttcttac taaatttgtg 180
aatctgggca ttcatcaagc a 201
<210> 38
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 38
aaatggtagc atacgtcact atccaacgaa tgacacagga tctggaaaat accctcccaa 60
aattgcatca ggaacaagta aacgttagca taaccccata wactttaagg gtttgcaaat 120
ccaacaatcc aagcattatt tgaaagttgt gggatgcaat atttcctgtg caggagcaaa 180
gggttaaaca aaattgctaa t 201
<210> 39
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 39
taaataactc catctcttat ttgtcaatcg taggccttcc agaacatcct caccatgagt 60
tgtataagag acagatgaag ggattcagtg gaatggtcac stttaaactg aaaggtaact 120
tggaaaatgc cacaaagttt ctccagacca tcaaagtgtt caccctggcc gaaagcctgg 180
gaggtttcga aagcctggtg g 201
<210> 40
<211> 201
<212> DNA
<213> Stichopus japonicus (Mare cucumis)
<400> 40
atcaacttgt ttcaaaatgg cctaccatta taaagccccg tagtagctta aatcatcttc 60
tggtcaacga catgttaagt ggcgatgcat tacaaagttc wccatccgtc tgttgcattt 120
tatgtctgta ttctgtccct acttcatcga cgccagggcg gaattggtac aaatttatag 180
tttgatgcca ctcatgtttc g 201
Claims (2)
1. A KASP labeled primer combination for identifying the hybrid stichopus japonicus in China and Russia is characterized in that the KASP labeled primer combination is shown in a table 1:
TABLE 110 KASP-tagged primer combinations for the detection of Russian-hybridized Stichopus japonicus
2. The use of the KASP marker primer combination of claim 1, wherein the specific application is the use of said primer combination to identify the population and individuals of the progeny stichopus japonicus from the russian cross or the stichopus japonicus from different batches, the specific method is as follows: extracting DNA of a stichopus japonicus sample by adopting an SDS method, carrying out SNP genotyping on the sample to be detected by using a KASP marker primer, wherein the judgment standard is as follows: the population detection rate reaches 70 percent, and is a Chinese and Russian hybrid stichopus japonicus population, and the population detection rate is the proportion of the number of individuals with at least 1 primer detected as a heterozygous type in the population to the total detected individuals; wherein the individual sample covered by at least 3 markers is filial stichopus japonicus or cross stichopus japonicus in different batches; the hybrid Chinese and Russian stichopus japonicus is a Water institute No.1 individual.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113151487A (en) * | 2021-02-09 | 2021-07-23 | 大连海洋大学 | Molecular identification marker primer combination for quantitative character of stichopus japonicus and thorn and application method thereof |
| CN114525362A (en) * | 2022-03-18 | 2022-05-24 | 大连海洋大学 | Primer combination for identifying Anyuan No. 1 stichopus japonicus population and application thereof |
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| CN101147473A (en) * | 2007-11-05 | 2008-03-26 | 大连水产学院 | Stichopus japonicus step-by-step hybrid breeding method |
| CN110724758A (en) * | 2019-11-27 | 2020-01-24 | 北京市农林科学院 | Method for identifying purity of Jingnongke 728 corn hybrid based on SNP marker |
| CN110777216A (en) * | 2019-11-27 | 2020-02-11 | 北京市农林科学院 | Method for identifying purity of Jingke waxy 2000 corn hybrid based on SNP marker |
| CN110872633A (en) * | 2019-11-27 | 2020-03-10 | 北京市农林科学院 | Method for identifying purity of Jingke 968 corn hybrid based on SNP marker |
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| CN101147473A (en) * | 2007-11-05 | 2008-03-26 | 大连水产学院 | Stichopus japonicus step-by-step hybrid breeding method |
| CN110724758A (en) * | 2019-11-27 | 2020-01-24 | 北京市农林科学院 | Method for identifying purity of Jingnongke 728 corn hybrid based on SNP marker |
| CN110777216A (en) * | 2019-11-27 | 2020-02-11 | 北京市农林科学院 | Method for identifying purity of Jingke waxy 2000 corn hybrid based on SNP marker |
| CN110872633A (en) * | 2019-11-27 | 2020-03-10 | 北京市农林科学院 | Method for identifying purity of Jingke 968 corn hybrid based on SNP marker |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113151487A (en) * | 2021-02-09 | 2021-07-23 | 大连海洋大学 | Molecular identification marker primer combination for quantitative character of stichopus japonicus and thorn and application method thereof |
| CN114525362A (en) * | 2022-03-18 | 2022-05-24 | 大连海洋大学 | Primer combination for identifying Anyuan No. 1 stichopus japonicus population and application thereof |
| CN114525362B (en) * | 2022-03-18 | 2023-05-23 | 大连海洋大学 | Primer combination for identifying Apostichopus japonicus population of Anyuan No. 1 and application thereof |
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