CN110438234A - A method of based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool - Google Patents
A method of based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool Download PDFInfo
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- CN110438234A CN110438234A CN201810409670.7A CN201810409670A CN110438234A CN 110438234 A CN110438234 A CN 110438234A CN 201810409670 A CN201810409670 A CN 201810409670A CN 110438234 A CN110438234 A CN 110438234A
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Abstract
The invention discloses a kind of methods based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool.The method of the present invention includes: the nucleotide in the genome for detect sheep to be measured in 11 groups of SNP site combinations at each SNP site, determines that the sheep to be measured corresponds to 11 genotype of 11 groups of SNP sites combination；Then according to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: if 11 genotype that the sheep to be measured corresponds to 11 groups of SNP sites combination meet the sheep known for its fine thick wool Reference genotype of respective sets SNP site combination, the sheep to be measured is or candidate is sheep known for its fine thick wool；If 11 genotype that the sheep to be measured corresponds to 11 groups of SNP sites combination meet the non-sheep known for its fine thick wool Reference genotype of respective sets SNP site combination, the sheep to be measured is or candidate is non-sheep known for its fine thick wool.The present invention is successfully obtained for identifying that sheep known for its fine thick wool and the SNP site of non-sheep known for its fine thick wool combine, and provides support for the identification of sheep known for its fine thick wool from now on, conservation and genetic breeding.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of side based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool
Method.
Background technique
Sheep known for its fine thick wool is the important herding resource in Ningxia, year the amount of delivering for sale up to 6,000,000, because of its distinctive flavor and the city meat Shen Shou
Field is praised highly, and is the considerable advantage brand in Ningxia.However, since the filial generation of the sheep known for its fine thick wool and other sheep verieties that occur recently emits
Sheep known for its fine thick wool sale is filled, huge damage is caused to sheep known for its fine thick wool brand.To stablize sheep known for its fine thick wool quality, sheep known for its fine thick wool brand image is renewed, sheep known for its fine thick wool is beaten
National premium brand is caused, we are educated by the identification to sheep known for its fine thick wool and non-sheep known for its fine thick wool for the mirror conservation of sheep known for its fine thick wool and heredity from now on
Kind lays the foundation.
Recently as the development of molecular biology, technique of gene detection also achieves considerable in biological species identification
Progress.Technique of gene detection can almost identify all biological species materials with it and be widely used in animals and plants object
Kind identification.Technique of gene detection identifies that animals and plants species are with fastest developing speed, the highest technological means of accuracy rate in species identification.
It is that the sequencing of full-length genome range is carried out to individual known to genome sequence that full-length genome, which resurveys sequence, with sequencing
Cost reduces and the species of known group sequence increase, and resurveys sequence and has become side maximally efficient in animals and plants breeding research
One of method.Sequenom SNP typing method is carried out by primer extension reaction principle, by sensitive PCR amplification and reliably
MALDI-TOF mass-spectrometric technique combines, and realizes Genotyping detection, this technology is particularly suitable for the knot to full-length genome research discovery
Fruit is verified.
Summary of the invention
The object of the present invention is to provide a kind of methods based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool.
In a first aspect, a kind of claimed method based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool.
Method provided by the present invention based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool can be following any:
The first, the method identified is combined based on 11 groups of SNP sites.This method specifically may include following steps: inspection
The nucleotide in the genome of sheep to be measured in 11 groups of SNP site combinations at each SNP site is surveyed, determines that the sheep to be measured corresponds to institute
State 11 genotype of 11 groups of SNP sites combination；Then according to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: if it is described to
It surveys sheep and corresponds to the sheep known for its fine thick wool standard base that 11 genotype that 11 groups of SNP sites combine meet the combination of respective sets SNP site
Because of type, then the sheep to be measured is or candidate is sheep known for its fine thick wool；If the sheep to be measured corresponds to 11 of 11 groups of SNP sites combination
Genotype meets the non-sheep known for its fine thick wool Reference genotype of respective sets SNP site combination, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool.
11 groups of SNP site groups are combined into following (a1)-(a11)；(a1) 11 groups of SNP sites shown in-(a11) combine institute
Corresponding non-sheep known for its fine thick wool Reference genotype and sheep known for its fine thick wool Reference genotype respectively are (b1)-(b11).I.e. SNP shown in (a1)
Point combination corresponding non-sheep known for its fine thick wool Reference genotype and sheep known for its fine thick wool Reference genotype are (b1)；(a2) SNP site shown in combines institute
Corresponding non-sheep known for its fine thick wool Reference genotype and sheep known for its fine thick wool Reference genotype are (b2)；... corresponding to the combination of SNP site shown in (a11)
Non- sheep known for its fine thick wool Reference genotype and sheep known for its fine thick wool Reference genotype be (b11).
(a1) contain 5 SNP sites, respectively CM001598.2:69009397, CM001602.2:49927473,
CM001603.2:38707088, CM001603.2:46619514 and CM001606.2:34435705；
(a2) contain 5 SNP sites, respectively CM001582.2:86006851, CM001598.2:69009397,
CM001600.2:45516740, CM001601.2:47737780 and CM001603.2:46619514；
(a3) contain 5 SNP sites, respectively CM001582.2:86006851, CM001583.2:42344562,
CM001584.2:11650210, CM001584.2:12200323 and CM001588.2:9423474；
(a4) contain 5 SNP sites, respectively CM001592.2:57058734, CM001595.2:48968164,
CM001598.2:69009397, CM001601.2:47737780 and CM001603.2:38707088；
(a5) contain 5 SNP sites, respectively CM001585.2:30868868, CM001589.2:75561002,
CM001590.2:56924859, CM001598.2:69009397 and CM001606.2:34435705；
(a6) contain 5 SNP sites, respectively CM001585.2:30868868, CM001588.2:84941340,
CM001598.2:45312379, CM001598.2:69009397 and CM001603.2:46619514；
(a7) contain 5 SNP sites, respectively CM001585.2:30868868, CM001588.2:84941340,
CM001594.2:45477903, CM001594.2:80961940 and CM001603.2:38707088；
(a8) contain 5 SNP sites, respectively CM001585.2:30868868, CM001588.2:84941340,
CM001593.2:1397868, CM001595.2:14087769 and CM001602.2:49927473；
(a9) contain 5 SNP sites, respectively CM001585.2:30868868, CM001588.2:84941340,
CM001592.2:34421625, CM001592.2:53664048 and CM001595.2:48968164；
(a10) contain 5 SNP sites, respectively CM001584.2:79082958, CM001584.2:157466635,
CM001601.2:47737780, CM001602.2:49927473 and CM001603.2:38707088；
(a11) contain 5 SNP sites, respectively CM001584.2:79082958, CM001585.2:5250467,
CM001588.2:84941340, CM001595.2:43390933 and CM001595.2:48968164；
The physical location of each SNP site in 11 groups of SNP sites combination is based on sheep known for its fine thick wool full-length genome standard sequence
Determination is compared, the version number of the sheep known for its fine thick wool full-length genome standard sequence is Ovis.Aries V4.0；Totally 29 SNP sites, tool
Body is as follows:
CM001582.2:86006851 is located on No. 1 chromosome the 86006851st, and deoxynucleotide is G or T；
CM001583.2:42344562 is located on No. 2 chromosomes the 42344562nd, and deoxynucleotide is T or C；
CM001584.2:11650210 is located on No. 3 chromosomes the 11650210th, and deoxynucleotide is T or C；
CM001584.2:12200323 is located on No. 3 chromosomes the 12200323rd, and deoxynucleotide is C or G；
CM001584.2:79082958 is located on No. 3 chromosomes the 79082958th, and deoxynucleotide is A or G；
CM001584.2:157466635 is located on No. 3 chromosomes the 157466635th, and deoxynucleotide is C or T；
CM001585.2:5250467 is located on No. 4 chromosomes the 5250467th, and deoxynucleotide is C or G；
CM001585.2:30868868 is located on No. 4 chromosomes the 30868868th, and deoxynucleotide is T or C；
CM001588.2:9423474 is located on No. 7 chromosomes the 9423474th, and deoxynucleotide is A or C；
CM001588.2:84941340 is located on No. 7 chromosomes the 84941340th, and deoxynucleotide is A or C；
CM001589.2:75561002 is located on No. 8 chromosomes the 75561002nd, and deoxynucleotide is C or G；
CM001590.2:56924859 is located on No. 9 chromosomes the 56924859th, and deoxynucleotide is A or G；
CM001592.2:34421625 is located on No. 11 chromosomes the 34421625th, and deoxynucleotide is T or C；
CM001592.2:53664048 is located on No. 11 chromosomes the 53664048th, and deoxynucleotide is A or G；
CM001592.2:57058734 is located on No. 11 chromosomes the 57058734th, and deoxynucleotide is C or G；
CM001593.2:1397868 is located on No. 12 chromosomes the 1397868th, and deoxynucleotide is A or G；
CM001594.2:45477903 is located on No. 13 chromosomes the 45477903rd, and deoxynucleotide is A or G；
CM001594.2:80961940 is located on No. 13 chromosomes the 80961940th, and deoxynucleotide is A or C；
CM001595.2:14087769 is located on No. 14 chromosomes the 14087769th, and deoxynucleotide is A or T；
CM001595.2:43390933 is located on No. 14 chromosomes the 43390933rd, and deoxynucleotide is A or G；
CM001595.2:48968164 is located on No. 14 chromosomes the 48968164th, and deoxynucleotide is T or G；
CM001598.2:45312379 is located on No. 17 chromosomes the 45312379th, and deoxynucleotide is T or C；
CM001598.2:69009397 is located on No. 17 chromosomes the 69009397th, and deoxynucleotide is A or C；
CM001600.2:45516740 is located on No. 19 chromosomes the 45516740th, and deoxynucleotide is A or C；
CM001601.2:47737780 is located on No. 20 chromosomes the 47737780th, and deoxynucleotide is A or G；
CM001602.2:49927473 is located on No. 21 chromosomes the 49927473rd, and deoxynucleotide is T or C；
CM001603.2:38707088 is located on No. 22 chromosomes the 38707088th, and deoxynucleotide is T or C；
CM001603.2:46619514 is located on No. 22 chromosomes the 46619514th, and deoxynucleotide is A or G；
CM001606.2:34435705 is located on No. 25 chromosomes the 34435705th, and deoxynucleotide is C or G；
Certainly, the version number of the used sheep known for its fine thick wool full-length genome standard sequence is not limited in the actual operation process
This is a kind of by Ovis.Aries V4.0, no matter using the version number of sheep known for its fine thick wool full-length genome standard sequence described in which kind of, as long as being related to
SNP site it is identical as 29 SNP sites of the invention described above, each fall within protection scope of the present invention.
(b1) non-sheep known for its fine thick wool Reference genotype is following any: CC_TT_CC_AA_CG；CC_TC_CC_AA_GG；CC_TT_
CC_AA_GG；CC_TC_TC_AA_GG；AC_TT_CC_AA_GG；AC_TT_CC_AA_CG；CC_CC_CC_AA_CG；AC_CC_
TC_AA_GG；AC_CC_CC_AA_CG；CC_CC_CC_AA_GG；CC_TC_TC_AA_CG；CC_TT_TC_AA_GG；CC_TC_
CC_AA_CG；CC_TT_TC_AA_CG；Sheep known for its fine thick wool Reference genotype is following any: CC_CC_TT_AA_CG；AA_CC_CC_AA_
CG；CC_TC_TT_AA_CG；CC_CC_CC_GA_CC；AC_TC_TC_AA_CG；AC_CC_TC_AA_CG；CC_CC_TC_AA_
CC；AC_TC_CC_AA_GG；AA_CC_TC_GA_CG；CC_CC_CC_GG_CG；AA_CC_CC_GG_CC；AC_CC_TC_GA_
GG；AA_CC_TC_AA_CC；CC_CC_TT_GA_GG；AC_CC_CC_AA_CC；AC_CC_TC_GA_CC.
(b2) non-sheep known for its fine thick wool Reference genotype is following any: TT_CC_CC_GG_AA；TT_AC_CC_GG_AA；TT_CC_
CC_AG_AA；TT_AC_AC_AG_AA；TT_CC_AC_AG_AA；TT_CC_AC_GG_AA；TT_CC_CC_AA_AA；Sheep known for its fine thick wool standard
Genotype is following any: TT_AA_AC_AG_GA；TT_CC_AA_AG_AA；GT_AC_CC_AG_AA；TT_AC_AC_GG_AA；
GG_AC_AC_GG_GA；TT_AC_CC_AG_AA；GT_AC_AC_AG_AA；GG_CC_CC_AG_GG；GG_AC_AC_AA_GA；
TT_CC_AC_GG_GA；GT_AA_AC_AG_AA；GG_CC_AA_AA_GA；GT_AC_AC_AG_GA；TT_AC_AC_AA_GA；
TT_CC_AC_AG_GA。
(b3) non-sheep known for its fine thick wool Reference genotype is following any: TT_TC_TT_CG_AC；TT_TT_TT_GG_AC；TT_TC_
TT_GG_CC；TT_CC_CT_GG_CC；TT_TT_CT_GG_CC；TT_TT_TT_GG_CC；TT_CC_TT_GG_CC；TT_TT_
TT_CG_CC；TT_TC_CT_GG_CC；TT_TC_TT_CG_CC；TT_TC_TT_GG_AC；GT_TT_TT_GG_CC；Sheep known for its fine thick wool standard
Genotype is following any: TT_CC_CT_CG_CC；TT_TC_CC_GG_CC；TT_TT_CT_CC_CC；GT_CC_CT_CG_AC；
GG_CC_TT_GG_AC；GT_CC_TT_GG_CC；TT_CC_TT_CG_AC；GG_TT_CC_CC_AA；TT_CC_CT_GG_AC；
TT_CC_CC_CG_AA；GT_CC_CT_CG_CC.
(b4) non-sheep known for its fine thick wool Reference genotype is following any: CC_GT_CC_GG_CC；GC_TT_CC_GG_CC；GG_TT_
CC_AG_TC；GG_GT_AC_GG_CC；GC_GT_CC_GG_CC；GC_TT_CC_AG_TC；CC_GT_AC_AG_CC；GC_TT_
CC_GG_TC；GG_TT_AC_GG_TC；GG_TT_CC_GG_CC；CC_TT_CC_AG_CC；CC_TT_AC_GG_CC；GC_TT_
CC_AG_CC；CC_TT_CC_GG_CC；CC_TT_CC_GG_TC；GC_GT_CC_AA_TC；GG_TT_CC_AG_CC；GG_GT_
CC_GG_CC；GC_GT_CC_GG_TC；Sheep known for its fine thick wool Reference genotype is following any: CC_GT_AA_AG_TC；GC_GT_AC_GG_
CC；GC_GT_AC_AG_TC；CC_GT_AA_AA_CC；GC_GG_AA_AG_TC；CC_GT_AC_AG_TT；CC_GG_CC_AA_
CC；GC_GG_AC_GG_TC；CC_TT_AC_GG_TT；CC_GT_AC_AG_TC；CC_GG_CC_AG_TC；CC_GG_AC_AG_
TC；CC_TT_CC_AG_TT；CC_GT_CC_AG_CC；CC_GG_AA_AG_CC；CC_TT_AC_AA_TC；CC_GG_AA_AG_
TC；CC_GT_CC_AA_TT；CC_GG_AC_AG_TT；CC_GT_AC_AA_CC；CC_GT_AC_AA_TC；CC_TT_AC_AG_
CC。
(b5) non-sheep known for its fine thick wool Reference genotype is following any: CC_GG_AA_CC_CG；CC_GG_GA_AC_GG；CC_GG_
GA_CC_GG；CC_CC_GA_CC_CG；CC_CC_AA_CC_GG；TC_GG_GG_AC_GG；CC_CG_GG_CC_CG；CC_CG_
GA_CC_GG；TC_CG_GA_AC_CG；CC_CG_GA_CC_CG；TC_CG_GA_AC_GG；CC_CG_GG_CC_GG；TC_CG_
AA_CC_GG；CC_CG_AA_CC_GG；CC_GG_GG_CC_GG；CC_GG_GA_AC_CG；TC_GG_GG_CC_GG；TC_GG_
GG_CC_CG；TC_GG_GA_CC_CG；TC_CG_GG_CC_CG；TC_GG_GA_CC_GG；Sheep known for its fine thick wool Reference genotype is following appoints
One: TC_CG_GA_AC_CC；TC_CC_GA_CC_CG；TT_CG_AA_AA_CG；CC_CC_AA_CC_CC；TT_CG_GA_AC_
CG；TC_CC_GA_AC_CG；TT_CC_AA_CC_CC；TC_CG_AA_CC_CG；TC_CG_GA_CC_CG；TC_GG_GA_AC_
GG；CC_CC_AA_CC_CG；TT_CG_GA_AA_CC；CC_CC_AA_AA_GG；TT_CC_GA_AC_CG；CC_CC_GA_AC_
CC。
(b6) non-sheep known for its fine thick wool Reference genotype is following any: CC_CA_CC_CC_AA；TC_AA_TC_CC_AA；CC_AA_
CC_AC_AA；CC_CA_TC_CC_AA；CC_CA_TT_CC_AA；TC_CA_CC_AC_AA；CC_AA_CC_CC_AA；CC_AA_
TC_CC_AA；TC_AA_CC_CC_AA；TC_CC_TC_CC_AA；CC_AA_TT_CC_AA；CC_CC_TC_CC_AA；TC_CA_
CC_CC_AA；Sheep known for its fine thick wool Reference genotype is following any: TC_CA_TC_AA_GA；TC_AA_TT_CC_AA；TT_CC_TC_AA_
AA；TT_AA_TT_AC_GA；TC_CA_TC_CC_AA；TC_AA_TC_AC_GA；TC_CC_TC_AC_AA；TC_AA_TT_AC_
AA；TT_CC_CC_CC_AA；TC_AA_TC_AC_AA；TC_CA_TT_AC_GA；CC_CA_TT_CC_GA；TT_CC_TT_AA_
AA；TC_CC_TC_CC_GA；TT_CC_TC_AC_GA；CC_CA_TT_AC_AA.
(b7) non-sheep known for its fine thick wool Reference genotype is following any: CC_CA_GG_AA_CC；TC_AA_GG_CA_CC；CC_CA_
GG_AA_TC；TC_CA_GG_AA_CC；CC_AA_GG_CA_CC；TC_CA_GG_CA_CC；TC_AA_GG_CA_TC；CC_AA_
GG_CA_TC；CC_CA_GG_CC_CC；CC_AA_GG_AA_CC；CC_CA_AG_AA_TC；TC_CC_GG_CA_CC；TC_AA_
GG_AA_TC；TC_CA_AG_AA_CC；CC_CC_GG_AA_CC；TC_AA_AG_AA_CC；Sheep known for its fine thick wool Reference genotype is following appoints
One: TC_CA_AG_AA_TC；TC_AA_AG_AA_TC；TT_AA_GG_AA_TC；TT_CC_AG_AA_CC；TT_CC_GG_AA_
TC；TT_AA_AG_AA_TC；TC_CA_AA_AA_TT；TC_CA_AA_AA_CC；TC_CC_AG_AA_TC；TC_CA_GG_AA_
TC；TC_CA_GG_AA_TT；TT_CC_AG_AA_TC；TC_CC_AG_AA_CC；CC_CA_AG_AA_CC.
(b8) non-sheep known for its fine thick wool Reference genotype is following any: CC_CA_GG_AA_TT；CC_AA_GG_AA_TC；CC_CA_
AG_AA_TT；CC_CA_GG_TT_TC；CC_CA_AG_AA_TC；TC_CA_AG_TA_TT；CC_AA_AG_TT_CC；CC_AA_
AG_TA_TC；CC_AA_AA_TA_CC；TC_AA_AG_TT_CC；CC_AA_GG_TT_TC；TC_AA_GG_AA_TT；CC_AA_
GG_TT_CC；CC_AA_AG_TA_CC；TC_AA_AG_TT_TC；CC_CA_AG_TT_CC；CC_AA_AG_TT_TC；CC_AA_
AG_AA_CC；CC_AA_GG_AA_CC；CC_CA_GG_TA_TC；TC_CC_GG_AA_TT；TC_AA_GG_TA_CC；TC_AA_
AG_AA_TC；CC_AA_GG_TT_TT；CC_AA_GG_TA_TT；CC_AA_AA_TA_TC；CC_CA_GG_TT_TT；CC_CC_
GG_TA_CC；TC_AA_GG_TA_TC；TC_CA_GG_TT_TC；CC_AA_GG_TA_TC；TC_AA_AG_AA_TT；Sheep known for its fine thick wool standard
Genotype is following any: TC_CA_AA_AA_TC；TC_CA_AG_AA_CC；TC_AA_AG_TA_CC；TT_CC_AA_AA_TC；
TT_AA_AA_AA_CC；TC_AA_AG_TA_TC；TC_CC_AG_AA_CC；TC_CA_GG_AA_CC；TC_CA_AG_AA_TC；
TC_AA_AG_AA_CC；TT_CC_GG_AA_CC；TC_CC_AA_AA_CC；CC_CA_AA_AA_TC；TT_CC_AA_AA_CC；
TT_AA_GG_AA_CC；CC_CA_GG_AA_CC.
(b9) non-sheep known for its fine thick wool Reference genotype is following any: CC_CA_TT_GG_GT；TC_AA_TT_GG_GT；CC_AA_
TT_AG_GT；TC_CA_TT_GG_GT；CC_AA_TT_GG_GT；CC_AA_CT_GG_TT；CC_AA_TT_GG_TT；TC_AA_
TT_AG_GT；TC_AA_TT_GG_TT；CC_CA_TT_GG_TT；CC_CA_TT_AG_TT；TC_CC_TT_GG_TT；CC_CC_
TT_AG_GT；TC_CA_TT_AG_TT；Sheep known for its fine thick wool Reference genotype is following any: TC_CA_TT_AG_GT；TT_AA_CT_AG_
GT；TT_CC_TT_AG_GG；TC_AA_TT_AG_TT；TC_AA_TT_GG_GG；TT_CC_CT_AG_GG；TC_CA_TT_GG_
TT；TC_CA_CT_GG_GT；CC_CA_TT_AG_GG；TT_CC_CT_AA_GG；TC_CC_CT_AG_GG；TC_CC_CT_AG_
GT；TT_AA_TT_AA_TT；TT_CC_TT_AA_GT；CC_CA_CT_AG_TT.
(b10) non-sheep known for its fine thick wool Reference genotype is following any: AA_CC_GG_TT_CC；AA_CT_GG_TT_CC；GA_CT_
GG_TT_CC；GA_CC_AG_TC_TC；GA_CC_AG_CC_CC；GA_CT_GG_TC_TC；GA_CC_GG_TC_CC；AA_CT_
GG_CC_CC；GA_CT_AG_CC_CC；AA_CT_AG_CC_CC；GA_TT_GG_CC_CC；AA_CC_AG_CC_CC；AA_CT_
AG_TC_TC；AA_CT_GG_TC_TC；GG_TT_GG_TT_CC；GA_TT_AA_TT_TC；GG_CC_AG_TC_CC；GA_CC_
AG_TT_CC；GA_CT_AG_TC_CC；AA_CC_GG_CC_CC；GA_TT_GG_TC_CC；AA_CC_GG_TC_CC；GA_CT_
GG_TC_CC；AA_CC_GG_TT_TC；Sheep known for its fine thick wool Reference genotype is following any: GA_CT_AG_TC_TC；AA_TT_GG_CC_
CC；GA_CT_AG_CC_TT；GG_TT_AG_TC_TT；GG_TT_GG_TC_TT；GG_CT_GG_TC_CC；GG_CT_AA_CC_
CC；GG_CT_GG_TC_TC；GG_CT_GG_TC_TT；AA_TT_AG_CC_TC；GA_CT_GG_CC_TC；GA_TT_AG_CC_
TC；AA_CT_AA_CC_TC；AA_CT_AG_TC_CC；GG_TT_AG_CC_TC；GG_TT_AG_CC_CC；GG_CT_AG_CC_
CC；GG_CT_AA_CC_TC；GG_TT_GG_TC_TC；GG_TT_AA_CC_CC；GA_TT_AA_CC_TC；GA_TT_AG_CC_
CC。
(b11) non-sheep known for its fine thick wool Reference genotype is following any: AA_GC_CA_AG_GT；GA_GC_AA_AG_GT；GG_GC_
AA_GG_GT；AA_CC_CA_AG_TT；GA_GC_CA_AG_TT；AA_CC_CA_GG_GT；GA_GC_AA_GG_GT；GA_CC_
AA_AA_GT；GA_CC_AA_AG_TT；AA_GG_AA_AG_TT；GA_CC_AA_GG_TT；AA_GG_AA_GG_TT；AA_CC_
AA_GG_TT；GA_GC_AA_GG_TT；AA_GC_AA_GG_TT；AA_GC_CA_AG_TT；AA_CC_AA_AG_TT；GA_GC_
AA_AG_TT；AA_GC_AA_AG_TT；AA_CC_CA_GG_TT；GA_CC_CC_AG_TT；AA_CC_AA_AA_TT；GG_CC_
AA_GG_TT；GA_CC_CA_AG_GT；AA_CC_CC_AG_GT；GA_CC_AA_AA_TT；AA_CC_AA_GG_GT；Sheep known for its fine thick wool standard
Genotype is following any: AA_GC_AA_GG_GT；AA_GC_CC_AG_GT；GA_GC_CA_AA_GT；GA_GG_CA_AG_TT；
GA_GG_AA_AG_GG；AA_GG_CC_AG_GG；GG_GC_CC_AA_GT；GG_GC_CA_AG_GT；GG_GC_CC_AA_GG；
GG_GC_CA_AA_TT；GA_GC_CA_AG_GG；GG_GC_CC_AG_GG；GG_CC_CC_AA_GG；GG_GC_CC_AG_GT；
GA_GC_CA_AG_GT；GA_GC_CC_AG_GT；AA_GG_CC_GG_GT.
Genotype shown in (b1)-(b11) is expressed as X above1X2_X3X4_X5X6_X7X8_X9X10, wherein X1To X10
Indicate A or C or T or G, X1X2(such as AA indicates the SNP site to nucleotide in expression SNP site combination at first SNP site
The nucleotide at place is the homozygous of A；As AT indicates that the nucleotide at the SNP site is the heterozygous of A and T), X3X4Indicate the SNP
Nucleotide ... ... in Sites Combination at second SNP site, X9X10It indicates in SNP site combination at the 5th SNP site
Nucleotide.
Second, the method identified is combined based on 1 group of SNP site.This method is concretely following any:
(1) include the following steps: to detect each in the combination of SNP site shown in (a1) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: such as
The genotype of sheep to be measured described in fruit meets the sheep known for its fine thick wool Reference genotype in (b1) described previously, then the sheep to be measured is or candidate is
Sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b1) described previously, the sheep to be measured
For or candidate be non-sheep known for its fine thick wool.
(2) include the following steps: to detect each in the combination of SNP site shown in (a2) described described in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: such as
The genotype of sheep to be measured described in fruit meets the sheep known for its fine thick wool Reference genotype in (b2), then the sheep to be measured is or candidate is
Sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b2), the sheep to be measured
For or candidate be non-sheep known for its fine thick wool.
(3) include the following steps: to detect each in the combination of SNP site shown in (a3) described described in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: such as
The genotype of sheep to be measured described in fruit meets the sheep known for its fine thick wool Reference genotype in (b3), then the sheep to be measured is or candidate is
Sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b3), the sheep to be measured
For or candidate be non-sheep known for its fine thick wool.
(4) include the following steps: to detect each in the combination of SNP site shown in (a4) described described in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: such as
The genotype of sheep to be measured described in fruit meets the sheep known for its fine thick wool Reference genotype in (b4), then the sheep to be measured is or candidate is
Sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b4) described previously, the sheep to be measured
For or candidate be non-sheep known for its fine thick wool.
(5) include the following steps: to detect each in the combination of SNP site shown in (a5) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: such as
The genotype of sheep to be measured described in fruit meets the sheep known for its fine thick wool Reference genotype in (b5) described previously, then the sheep to be measured is or candidate is
Sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b5) described previously, the sheep to be measured
For or candidate be non-sheep known for its fine thick wool.
(6) include the following steps: to detect each in the combination of SNP site shown in (a6) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: such as
The genotype of sheep to be measured described in fruit meets the sheep known for its fine thick wool Reference genotype in (b6) described previously, then the sheep to be measured is or candidate is
Sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b6) described previously, the sheep to be measured
For or candidate be non-sheep known for its fine thick wool.
(7) include the following steps: to detect each in the combination of SNP site shown in (a7) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: such as
The genotype of sheep to be measured described in fruit meets the sheep known for its fine thick wool Reference genotype in (b7) described previously, then the sheep to be measured is or candidate is
Sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b7) described previously, the sheep to be measured
For or candidate be non-sheep known for its fine thick wool.
(8) include the following steps: to detect each in the combination of SNP site shown in (a8) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: such as
The genotype of sheep to be measured described in fruit meets the sheep known for its fine thick wool Reference genotype in (b8) described previously, then the sheep to be measured is or candidate is
Sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b8) described previously, the sheep to be measured
For or candidate be non-sheep known for its fine thick wool.
(9) include the following steps: to detect each in the combination of SNP site shown in (a9) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: such as
The genotype of sheep to be measured described in fruit meets the sheep known for its fine thick wool Reference genotype in (b9) described previously, then the sheep to be measured is or candidate is
Sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b9) described previously, the sheep to be measured
For or candidate be non-sheep known for its fine thick wool.
(10) include the following steps: to detect in the combination of SNP site shown in (a10) described previously in the genome of sheep to be measured
Nucleotide at each SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows:
If the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b10) described previously, the sheep to be measured is or waits
It is selected as sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b10) described previously, it is described to
Survey sheep is or candidate is non-sheep known for its fine thick wool.
(11) include the following steps: to detect in the combination of SNP site shown in (a11) described previously in the genome of sheep to be measured
Nucleotide at each SNP site determines the genotype of the sheep to be measured；According to determining whether the sheep to be measured is sheep known for its fine thick wool as follows:
If the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b11) described previously, the sheep to be measured is or waits
It is selected as sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool Reference genotype in (b11) described previously, it is described to
Survey sheep is or candidate is non-sheep known for its fine thick wool.
Second aspect, a kind of claimed method based on Genotyping identification sheep known for its fine thick wool.
Method provided by the present invention based on Genotyping identification sheep known for its fine thick wool can be following any:
The first, the method identified is combined based on 11 groups of SNP sites.This method may include following steps: detection to
The nucleotide in the genome of sheep in 11 groups of SNP site combinations at each SNP site is surveyed, determines that the sheep to be measured corresponds to described 11
11 genotype of group SNP site combination；If the sheep to be measured corresponds to 11 genotype of 11 groups of SNP sites combination
Meet the sheep known for its fine thick wool Reference genotype of respective sets SNP site combination, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
Wherein, 11 groups of SNP site groups are combined into previously described (a1)-(a11)；(a1) 11 groups shown in-(a11)
The corresponding sheep known for its fine thick wool Reference genotype of SNP site combination respectively is the sheep known for its fine thick wool standard base in previously described (b1)-(b11)
Because of type.
Second, the method identified is combined based on 1 group of SNP site.This method is concretely following any:
(1) include the following steps: to detect each in the combination of SNP site shown in (a1) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b1) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(2) include the following steps: to detect each in the combination of SNP site shown in (a2) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b2) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(3) include the following steps: to detect each in the combination of SNP site shown in (a3) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b3) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(4) include the following steps: to detect each in the combination of SNP site shown in (a4) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b4) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(5) include the following steps: to detect each in the combination of SNP site shown in (a5) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b5) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(6) include the following steps: to detect each in the combination of SNP site shown in (a6) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b6) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(7) include the following steps: to detect each in the combination of SNP site shown in (a7) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b7) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(8) include the following steps: to detect each in the combination of SNP site shown in (a8) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b8) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(9) include the following steps: to detect each in the combination of SNP site shown in (a9) described previously in the genome of sheep to be measured
Nucleotide at SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b9) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(10) include the following steps: to detect in the combination of SNP site shown in (a10) described previously in the genome of sheep to be measured
Nucleotide at each SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b10) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
(11) include the following steps: to detect in the combination of SNP site shown in (a11) described previously in the genome of sheep to be measured
Nucleotide at each SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets described previously
(b11) the sheep known for its fine thick wool Reference genotype in, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
In the method described in first aspect and second aspect, the sheep to be measured concretely (such as cover by sheep known for its fine thick wool or non-sheep known for its fine thick wool
Ancient sheep).
In the method described in first aspect and second aspect, detect in the genome of the sheep to be measured at SNP site
The specific method of nucleotide can be full-length genome weight PCR sequencing PCR.Further, for carrying out the full-length genome weight PCR sequencing PCR
DNA sample is taken from the blood or ear tissue of the sheep to be measured.
Following several applications are also claimed in the third aspect, the present invention.
The first, for detecting the nucleosides in sheep genome in previously described 11 groups of SNP sites combination at each SNP site
The substance (or substance for detecting the nucleotide in sheep genome at previously described 29 SNP sites) of acid is on identification beach
Application in sheep and/or non-sheep known for its fine thick wool, or preparing the application in the kit for identifying sheep known for its fine thick wool and/or non-sheep known for its fine thick wool.
Second, for detecting in sheep genome each SNP of any one group in the combination of previously described 11 groups of SNP sites
The identification sheep known for its fine thick wool of nucleotide at point and/or the application in non-sheep known for its fine thick wool, or in preparation for identifying sheep known for its fine thick wool and/or non-beach
Application in the kit of sheep.
In above two application, the substance be concretely directed to PCR primer designed by SNP site to be detected and/
Or sequencing primer.
Further, the primer for detecting the CM001582.2:86006851 is SEQ ID No.1 and SEQ ID
Two single stranded DNAs shown in No.2；Primer for detecting the CM001583.2:42344562 is SEQ ID No.4 and SEQ
Two single stranded DNAs shown in ID No.5；For detect the CM001584.2:11650210 primer be SEQ ID No.7 and
Two single stranded DNAs shown in SEQ ID No.8；Primer for detecting the CM001584.2:12200323 is SEQ ID
Two single stranded DNAs shown in No.10 and SEQ ID No.11；Primer for detecting the CM001584.2:79082958 is SEQ
Two single stranded DNAs shown in ID No.13 and SEQ ID No.14；For detecting the primer of the CM001584.2:157466635
For two single stranded DNAs shown in SEQ ID No.16 and SEQ ID No.17；For detecting the CM001585.2:5250467's
Primer is two single stranded DNAs shown in SEQ ID No.19 and SEQ ID No.20；For detecting the CM001585.2:
30868868 primer is two single stranded DNAs shown in SEQ ID No.22 and SEQ ID No.23；It is described for detecting
The primer of CM001588.2:9423474 is two single stranded DNAs shown in SEQ ID No.25 and SEQ ID No.26；For detecting
The primer of the CM001588.2:84941340 is two single stranded DNAs shown in SEQ ID No.28 and SEQ ID No.29；For
The primer for detecting the CM001589.2:75561002 is two single stranded DNAs shown in SEQ ID No.31 and SEQ ID No.32；
It is single-stranded for two shown in SEQ ID No.34 and SEQ ID No.35 for detecting the primer of the CM001590.2:56924859
DNA；Primer for detecting the CM001592.2:34421625 is two shown in SEQ ID No.37 and SEQ ID No.38
Single stranded DNA；Primer for detecting the CM001592.2:53664048 is shown in SEQ ID No.40 and SEQ ID No.41
Two single stranded DNAs；Primer for detecting the CM001592.2:57058734 is SEQ ID No.43 and SEQ ID No.44
Shown two single stranded DNAs；Primer for detecting the CM001593.2:1397868 is SEQ ID No.46 and SEQ ID
Two single stranded DNAs shown in No.47；Primer for detecting the CM001594.2:45477903 is SEQ ID No.49 and SEQ
Two single stranded DNAs shown in ID No.50；For detect the CM001594.2:80961940 primer be SEQ ID No.52 and
Two single stranded DNAs shown in SEQ ID No.53；Primer for detecting the CM001595.2:14087769 is SEQ ID
Two single stranded DNAs shown in No.55 and SEQ ID No.56；Primer for detecting the CM001595.2:43390933 is SEQ
Two single stranded DNAs shown in ID No.58 and SEQ ID No.59；Primer for detecting the CM001595.2:48968164 is
Two single stranded DNAs shown in SEQ ID No.61 and SEQ ID No.62；For detecting drawing for the CM001598.2:45312379
Object is two single stranded DNAs shown in SEQ ID No.64 and SEQ ID No.65；For detecting the CM001598.2:69009397
Primer be SEQ ID No.67 and SEQ ID No.68 shown in two single stranded DNAs；For detecting the CM001600.2:
45516740 primer is two single stranded DNAs shown in SEQ ID No.70 and SEQ ID No.71；It is described for detecting
The primer of CM001601.2:47737780 is two single stranded DNAs shown in SEQ ID No.73 and SEQ ID No.74；For detecting
The primer of the CM001602.2:49927473 is two single stranded DNAs shown in SEQ ID No.76 and SEQ ID No.77；For
The primer for detecting the CM001603.2:38707088 is two single stranded DNAs shown in SEQ ID No.79 and SEQ ID No.80；
It is single-stranded for two shown in SEQ ID No.82 and SEQ ID No.83 for detecting the primer of the CM001603.2:46619514
DNA；Primer for detecting the CM001606.2:34435705 is two shown in SEQ ID No.85 and SEQ ID No.86
Single stranded DNA.
Further, the primer for detecting the CM001582.2:86006851 further includes shown in SEQ ID No.3
Single stranded DNA；Primer for detecting the CM001583.2:42344562 further includes single-stranded shown in SEQ ID No.6
DNA；Primer for detecting the CM001584.2:11650210 further includes single stranded DNA shown in SEQ ID No.9；For
The primer for detecting the CM001584.2:12200323 further includes single stranded DNA shown in SEQ ID No.12；It is described for detecting
The primer of CM001584.2:79082958 further includes single stranded DNA shown in SEQ ID No.15；It is described for detecting
The primer of CM001584.2:157466635 further includes single stranded DNA shown in SEQ ID No.18；It is described for detecting
The primer of CM001585.2:5250467 further includes single stranded DNA shown in SEQ ID No.21；It is described for detecting
The primer of CM001585.2:30868868 further includes single stranded DNA shown in SEQ ID No.24；It is described for detecting
The primer of CM001588.2:9423474 further includes single stranded DNA shown in SEQ ID No.27；It is described for detecting
The primer of CM001588.2:84941340 further includes single stranded DNA shown in SEQ ID No.30；It is described for detecting
The primer of CM001589.2:75561002 further includes single stranded DNA shown in SEQ ID No.33；It is described for detecting
The primer of CM001590.2:56924859 further includes single stranded DNA shown in SEQ ID No.36；It is described for detecting
The primer of CM001592.2:34421625 further includes single stranded DNA shown in SEQ ID No.39；It is described for detecting
The primer of CM001592.2:53664048 further includes single stranded DNA shown in SEQ ID No.42；It is described for detecting
The primer of CM001592.2:57058734 further includes single stranded DNA shown in SEQ ID No.45；It is described for detecting
The primer of CM001593.2:1397868 further includes single stranded DNA shown in SEQ ID No.48；It is described for detecting
The primer of CM001594.2:45477903 further includes single stranded DNA shown in SEQ ID No.51；It is described for detecting
The primer of CM001594.2:80961940 further includes single stranded DNA shown in SEQ ID No.54；It is described for detecting
The primer of CM001595.2:14087769 further includes single stranded DNA shown in SEQ ID No.57；It is described for detecting
The primer of CM001595.2:43390933 further includes single stranded DNA shown in SEQ ID No.60；It is described for detecting
The primer of CM001595.2:48968164 further includes single stranded DNA shown in SEQ ID No.63；It is described for detecting
The primer of CM001598.2:45312379 further includes single stranded DNA shown in SEQ ID No.66；It is described for detecting
The primer of CM001598.2:69009397 further includes single stranded DNA shown in SEQ ID No.69；It is described for detecting
The primer of CM001600.2:45516740 further includes single stranded DNA shown in SEQ ID No.72；It is described for detecting
The primer of CM001601.2:47737780 further includes single stranded DNA shown in SEQ ID No.75；It is described for detecting
The primer of CM001602.2:49927473 further includes single stranded DNA shown in SEQ ID No.78；It is described for detecting
The primer of CM001603.2:38707088 further includes single stranded DNA shown in SEQ ID No.81；It is described for detecting
The primer of CM001603.2:46619514 further includes single stranded DNA shown in SEQ ID No.84；It is described for detecting
The primer of CM001606.2:34435705 further includes single stranded DNA shown in SEQ ID No.87.
The third, nucleotide in sheep genome in the combination of previously described 11 groups of SNP sites at each SNP site it is polymorphic
Property (or polymorphism of the nucleotide in sheep genome at previously described 29 SNP sites) in identification sheep known for its fine thick wool and/or non-beach
Application in sheep.
4th kind, the core in sheep genome in previously described 11 groups of SNP sites combination at any one group of each SNP site
Application of the polymorphism of thuja acid in identification sheep known for its fine thick wool and/or non-sheep known for its fine thick wool.
Fourth aspect, primer set shown in claimed following (B1) or (B2):
(B1) for detecting the nucleotide in sheep genome in previously described 11 groups of SNP sites combination at each SNP site
Primer set；
(B2) it is used to detect in sheep genome any one group in previously described 11 groups of SNP sites combination of each SNP site
The primer set of the nucleotide at place.
Wherein, for detecting the primer of each SNP site as mentioned before (see Table 2 for details).
The present invention resurveys sequence by carrying out full-length genome to 50 sheep known for its fine thick wools and 50 non-sheep known for its fine thick wools, obtains 29 sites, then
By R Analysis of programming, obtain 5 genotyping result genotype (identification rate is 85% or more).Then pass through 1000 portions of sheep known for its fine thick wools and 1000
The non-sheep known for its fine thick wool of part carries out sequenom verifying, is consistent as the result is shown with result is determined.The present invention is successfully obtained for identifying beach
Sheep combines with the SNP site of non-sheep known for its fine thick wool, this for exploitation sheep known for its fine thick wool cultivar identification chip, more the identification of sheep known for its fine thick wool from now on, conservation and
Genetic breeding provides support.
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1, full-length genome resurvey the 5 genotyping result genotype that sequence obtains identification sheep known for its fine thick wool and non-sheep known for its fine thick wool
For sample sheet: 50 sheep known for its fine thick wools (coming from Yanchi county Ningxia) and 50 non-sheep known for its fine thick wools (from Ningxia, Inner Mongol and other places).
So that the blood or ear tissue of examination sheep are sample, genomic DNA is extracted, the side of sequence is then resurveyed using full-length genome
Method obtains 29 SNP sites, then by R Analysis of programming, obtains 5 genotyping result genotype (table 1).
Wherein, SNP site selection method is as follows: marking the site of all value < -8 P first, then manually observation is
It is no to have continuous chromosomal region (linkage disequilibrium region).Continuum is just regarded as when at least more than 4 continuous sites.
Select principle in site are as follows: 1) select the most significant site in the site of those value < -8 P in continuum first.2) it secondly selects
Those P values < -8 are selected, but the most significant site not in the site of continuum；3) those biggish continuums are selected, still
But there is no the most significant site in the site of value < -8 P.
The method that full-length genome resurveys sequence is specific as follows: genomic DNA extracted, is interrupted at random using Covaris, electricity
The DNA fragmentation (0.2~5Kb) of length needed for swimming is recycled, in addition connector, carries out cluster and prepare (Illumina Hiseq
X10), finally Insert Fragment is carried out to resurvey sequence using the method for Paired-End (Illumina Hiseq X10).
The physical location of 29 SNP sites is to compare determination based on sheep known for its fine thick wool full-length genome standard sequence, and the sheep known for its fine thick wool is complete
The version number of genome standard sequence is Ovis.Aries V4.0；It is specific as follows:
CM001582.2:86006851 is located on No. 1 chromosome the 86006851st, and deoxynucleotide is G or T；
CM001583.2:42344562 is located on No. 2 chromosomes the 42344562nd, and deoxynucleotide is T or C；
CM001584.2:11650210 is located on No. 3 chromosomes the 11650210th, and deoxynucleotide is T or C；
CM001584.2:12200323 is located on No. 3 chromosomes the 12200323rd, and deoxynucleotide is C or G；
CM001584.2:79082958 is located on No. 3 chromosomes the 79082958th, and deoxynucleotide is A or G；
CM001584.2:157466635 is located on No. 3 chromosomes the 157466635th, and deoxynucleotide is C or T；
CM001585.2:5250467 is located on No. 4 chromosomes the 5250467th, and deoxynucleotide is C or G；
CM001585.2:30868868 is located on No. 4 chromosomes the 30868868th, and deoxynucleotide is T or C；
CM001588.2:9423474 is located on No. 7 chromosomes the 9423474th, and deoxynucleotide is A or C；
CM001588.2:84941340 is located on No. 7 chromosomes the 84941340th, and deoxynucleotide is A or C；
CM001589.2:75561002 is located on No. 8 chromosomes the 75561002nd, and deoxynucleotide is C or G；CM001590.2:
56924859 are located on No. 9 chromosomes the 56924859th, and deoxynucleotide is A or G；CM001592.2:34421625
In the 34421625th on No. 11 chromosomes, deoxynucleotide is T or C；CM001592.2:53664048 is located at No. 11 dyeing
The 53664048th on body, deoxynucleotide is A or G；CM001592.2:57058734 is located on No. 11 chromosomes the
57058734, deoxynucleotide is C or G；CM001593.2:1397868 is located on No. 12 chromosomes the 1397868th,
Its deoxynucleotide is A or G；CM001594.2:45477903 is located on No. 13 chromosomes the 45477903rd, deoxyribonucleoside
Acid is A or G；CM001594.2:80961940 is located on No. 13 chromosomes the 80961940th, and deoxynucleotide is A or C；
CM001595.2:14087769 is located on No. 14 chromosomes the 14087769th, and deoxynucleotide is A or T；
CM001595.2:43390933 is located on No. 14 chromosomes the 43390933rd, and deoxynucleotide is A or G；
CM001595.2:48968164 is located on No. 14 chromosomes the 48968164th, and deoxynucleotide is T or G；
CM001598.2:45312379 is located on No. 17 chromosomes the 45312379th, and deoxynucleotide is T or C；
CM001598.2:69009397 is located on No. 17 chromosomes the 69009397th, and deoxynucleotide is A or C；
CM001600.2:45516740 is located on No. 19 chromosomes the 45516740th, and deoxynucleotide is A or C；
CM001601.2:47737780 is located on No. 20 chromosomes the 47737780th, and deoxynucleotide is A or G；
CM001602.2:49927473 is located on No. 21 chromosomes the 49927473rd, and deoxynucleotide is T or C；
CM001603.2:38707088 is located on No. 22 chromosomes the 38707088th, and deoxynucleotide is T or C；
CM001603.2:46619514 is located on No. 22 chromosomes the 46619514th, and deoxynucleotide is A or G；
CM001606.2:34435705 is located on No. 25 chromosomes the 34435705th, and deoxynucleotide is C or G；
Table 1 identifies sheep known for its fine thick wool and non-5 parting loci gene type of sheep known for its fine thick wool
Note: the genotype of 5 SNP sites is represented as X in each group1X2_X3X4_X5X6_X7X8_X9X10, wherein X1To X10
Indicate A or C or T or G, X1X2(such as AA indicates the nucleosides at the SNP site to the nucleotide for indicating in the group at first SNP site
Acid is the homozygous of A；As AT indicates that the nucleotide at the SNP site is the heterozygous of A and T), X3X4Second is indicated in the group
Nucleotide ... ... at SNP site, X9X10Indicate the nucleotide in the group at the 5th SNP site.
Embodiment 2, the 5 genotyping result genotype for verifying sheep known for its fine thick wool and non-sheep known for its fine thick wool
Verifying sample: 1000 portions of sheep known for its fine thick wools and 1000 parts of non-sheep known for its fine thick wools.
To verify blood or ear tissue with sheep as sample, genomic DNA is extracted, Sequenom verifying is then carried out.
Wherein, Sequenom verifying specific steps are as follows:
1. design of primers
It is to be measured using Sequenom company Genotyping Tools and MassARRAY Assay Design software design
The PCR amplification primer and Single base extension primer of SNP site, and biotech firm is transferred to synthesize.
2.DNA quality inspection
Quantitative with spectrophotometer to experiment sample, agarose gel electrophoresis quality inspection, genome dna electrophoresis band is usually not
Less than 20kb.Concentration is adjusted to 50ng/ μ l by the DNA of quality inspection qualification, is transferred to 96 orifice plates, and -20 DEG C store for future use.
3. primer dilutes
(1) each SNP has three primers (table 2), and the number of this three primers is marked to the pipe lid in primer composite tube
On.
The primer information of each SNP of table 2
(2) Forward PCR primer, Reverse PCR primer are first centrifuged sky and get rid of Hou Jiashui, and amount of water is related to OD value
(primer pipe and design of primers table have mark), every 1OD add 36 μ l water.Concussion is added after being placed at room temperature for 30min after water to mix spare.
(3) calculation method when the Forward PCR primer, Reverse PCR primer of multiple SNP mix in each Well:
Forward PCR primer, the Reverse PCR primer of each SNP respectively takes 2.5 μ l amount of water (μ l)=500- tuple * 2.5*2
(tuple: SNP number in well)
4.PCR amplification
PCR amplification uses multiple PCR technique, carries out in 384 orifice plates, and each reaction system total volume is 5 μ l.
(1) PCR master mix solution is prepared in a new 2.0ml EP pipe, it is specific as shown in table 3.
3 PCR master mix solution of table
(2) will be spare in the PCR pipe for splitting 8 platoons after the concussion mixing of prepared PCR master mix solution, use 8
Channel sample injector, adjusting injection volume are 4 μ L, and PCR master mix liquid is added in each well of 384 orifice plates.It should
384 orifice plates are PCR reaction plate.
(3) 96 orifice plate of DNA sample prepared is taken out, using 8 channel sample injectors, adjusting injection volume is 1 μ L, is added
To corresponding 384PCR reaction plate, sealed membrane is sticked, sky is got rid of after concussion mixes.
(4) setting PCR reaction condition is as shown in table 4 in the PCR instrument of compatible 384 orifice plates.
4 PCR reaction condition of table
5.PCR product alkaline phosphatase treatment
(1) PCR after reaction, 384 reaction plates take out and sky get rid of.
(2) alkaline phosphatase treatment reaction solution is prepared, SAP Mix is specific as shown in table 5.
5 alkaline phosphatase treatment reaction solution SAP Mix of table
| SAP Mix | To each reaction (μ L) |
| Water | 1.53 |
| SAP Buffer(10x) | 0.17 |
| SAP enzyme (1.7U/ul) | 0.3 |
| Total volume | 2 |
(3) will be spare in the PCR pipe for splitting 8 platoons after the concussion mixing of prepared SAP mix solution, added using 8 channels
Sample device, adjusting injection volume are 2 μ L, and 384 hole PCR reaction plates are added in SAP Mix.Sealed membrane is sticked, sky is got rid of after concussion mixes.
Reaction system total volume is 7 μ l (wherein 5 μ l, SAP mixed liquor of PCR product, 2 μ l).
(4) 384 orifice plates are placed in the PCR instrument of compatible 384 orifice plates, setting PCR reaction condition is as shown in table 6.
6 PCR reaction condition of table
| Temperature (DEG C) | Time (minute) | Circulation |
| 37 | 40 | 1 |
| 85 | 5 | 1 |
| 4 | ∞ | 1 |
Start PCR instrument and carries out alkaline phosphatase treatment reaction.
6. Single base extension
(1) after alkaline phosphatase treatment, sky is got rid of after 384 reaction plates are taken out, and is carrying out single base extension,
9 μ l of reaction system total volume.
(2) preparation single base extension liquid, EXTEND Mix, as shown in table 7.(note: Extend primer Mix
It is had to Well number corresponding correct)
7 single base extension liquid EXTEND Mix of table
| EXTEND Mix | To each reaction (μ L) |
| Water | 0.619 |
| Extend primer Mix | 0.94 |
| iPLEX Buffer plus | 0.2 |
| iPLEX terminator | 0.2 |
| iPLEX enzyme | 0.041 |
| Total volume | 2 |
(3) will be spare in the PCR pipe for splitting 8 platoons after the concussion mixing of prepared EXTEND mix solution, use 8 channels
Sample injector, adjusting injection volume are 2 μ L, by EXTEND Mix 384 hole reaction plates of corresponding addition.For each reacting hole, single alkali
Base extension system includes 7 μ l and EXTEND Mix liquid of PCR product 2 μ l, total system 9ul after SAP processing.
(4) 384 orifice plates are placed in the PCR instrument of compatible 384 orifice plates, set PCR reaction condition, as shown in table 8.
8 PCR reaction condition of table
Start PCR instrument and carries out single base extension.
7. purifying resin
Reaction product plus 16 μ l water are diluted, carry out desalination using resin after dilution.
8. chip point sample
By the sample spot after desalting processing on sample target, spontaneous nucleation.
9. Mass Spectrometer Method
Upper machine carries out Mass Spectrometer Method, and collects data.
The results show that the identification rate of this method 85% or more (indicate: 100 sheep to be measured are identified in the method,
There are 85 or more can identify result), it is specific as shown in table 9.SNP site shown in sheep known for its fine thick wool and non-sheep known for its fine thick wool table 1 combines base
Because type is shown in table 1 within the scope of corresponding gene type, see Table 10 for details for ratio.
The identification rate of 9 the method for the present invention of table
10 sheep known for its fine thick wool of table and non-sheep known for its fine thick wool Sequenom SNP testing result (5 parting loci gene type proportion)
Note: the genotype representation method in table is the same as table 1.
The result shows that verifying the 5 genotyping result genes of sheep known for its fine thick wool and non-sheep known for its fine thick wool shown in the resulting table 1 of embodiment 1 shown in table 9
Type is accurate and reliable.The present invention is successfully obtained for identifying that sheep known for its fine thick wool and the SNP site of non-sheep known for its fine thick wool combine, be the identification of sheep known for its fine thick wool from now on,
Conservation and genetic breeding provide support.
<110>Ningxia Academy of Agri-Forestry Sciences's animal science research institute；Institute of Animal Sciences, Chinese Academy of Agricultural Sciences；Beijing
Kang Pusen Bioisystech Co., Ltd
<120>a kind of method based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool
<130> GNCLN180740
<160> 87
<170> PatentIn version 3.5
<210> 1
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<212> DNA
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acgttggatg acccacacca cacagcttg 29
<210> 2
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acgttggatg aagtcacata gtcctggtgc 30
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<400> 3
ctctggggct cagtgt 16
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acgttggatg ttatacttgg gcctcagcag 30
<210> 5
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<400> 5
acgttggatg actgaagaca gccaggtcac 30
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<400> 6
tctggcacca cagtggccac taga 24
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acgttggatg gggtaagtgt gaactccaag 30
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<400> 8
acgttggatg tctctcggtg atgtggttgg 30
<210> 9
<211> 15
<212> DNA
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<400> 9
actccaagag ggaca 15
<210> 10
<211> 30
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<400> 10
acgttggatg aaaggcctca gggattgatg 30
<210> 11
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<400> 11
acgttggatg aatcagctca gtgtcgctgg 30
<210> 12
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<400> 12
ggaatgcgac aggcagcttc 20
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<211> 30
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<400> 13
acgttggatg tgattcccac tccaatcagg 30
<210> 14
<211> 30
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<400> 14
acgttggatg tcatggcatg gttcagagtc 30
<210> 15
<211> 19
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<213> Artificial sequence
<400> 15
gacgcactat acggcacca 19
<210> 16
<211> 30
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<213> Artificial sequence
<400> 16
acgttggatg tgacaaggac taagcaaagc 30
<210> 17
<211> 30
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<400> 17
acgttggatg gatggcagcc ctagatcttc 30
<210> 18
<211> 18
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<400> 18
cgggtaaaga ttgcaggt 18
<210> 19
<211> 29
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<400> 19
acgttggatg tgcaggcgag aactcacag 29
<210> 20
<211> 30
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<400> 20
acgttggatg tggtctgccc cagagaaatg 30
<210> 21
<211> 20
<212> DNA
<213> Artificial sequence
<400> 21
cccccatggc cctgtgctgg 20
<210> 22
<211> 30
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<400> 22
acgttggatg ttctggttcc aactggtctg 30
<210> 23
<211> 30
<212> DNA
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<400> 23
acgttggatg ttgcagatac cgaaacctcc 30
<210> 24
<211> 17
<212> DNA
<213> Artificial sequence
<400> 24
ccaggtggaa gaaggta 17
<210> 25
<211> 30
<212> DNA
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<400> 25
acgttggatg tttccccaag gtcacatagc 30
<210> 26
<211> 30
<212> DNA
<213> Artificial sequence
<400> 26
acgttggatg caatacagca gaacaccgac 30
<210> 27
<211> 19
<212> DNA
<213> Artificial sequence
<400> 27
cccccgacgg acataactc 19
<210> 28
<211> 30
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<400> 28
acgttggatg tgggtcctgg ggagcagcaa 30
<210> 29
<211> 29
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<213> Artificial sequence
<400> 29
acgttggatg tctctgcact ctgcctctc 29
<210> 30
<211> 15
<212> DNA
<213> Artificial sequence
<400> 30
gagcagcaat gggcg 15
<210> 31
<211> 30
<212> DNA
<213> Artificial sequence
<400> 31
acgttggatg ggtcatgctc aatattcacc 30
<210> 32
<211> 30
<212> DNA
<213> Artificial sequence
<400> 32
acgttggatg aacctgggtc tcccatattg 30
<210> 33
<211> 17
<212> DNA
<213> Artificial sequence
<400> 33
ccatattgca ggcagac 17
<210> 34
<211> 30
<212> DNA
<213> Artificial sequence
<400> 34
acgttggatg tgattcccac tccaatcagg 30
<210> 35
<211> 30
<212> DNA
<213> Artificial sequence
<400> 35
acgttggatg tcatggcatg gttcagagtc 30
<210> 36
<211> 16
<212> DNA
<213> Artificial sequence
<400> 36
gcactatacg gcacca 16
<210> 37
<211> 30
<212> DNA
<213> Artificial sequence
<400> 37
acgttggatg tgacaaggac taagcaaagc 30
<210> 38
<211> 30
<212> DNA
<213> Artificial sequence
<400> 38
acgttggatg gatggcagcc ctagatcttc 30
<210> 39
<211> 25
<212> DNA
<213> Artificial sequence
<400> 39
tagatcttca tattttaggt tcctt 25
<210> 40
<211> 29
<212> DNA
<213> Artificial sequence
<400> 40
acgttggatg tgcaggcgag aactcacag 29
<210> 41
<211> 30
<212> DNA
<213> Artificial sequence
<400> 41
acgttggatg tggtctgccc cagagaaatg 30
<210> 42
<211> 19
<212> DNA
<213> Artificial sequence
<400> 42
cgaaatggcc ctgtgctgg 19
<210> 43
<211> 30
<212> DNA
<213> Artificial sequence
<400> 43
acgttggatg ttctggttcc aactggtctg 30
<210> 44
<211> 30
<212> DNA
<213> Artificial sequence
<400> 44
acgttggatg ttgcagatac cgaaacctcc 30
<210> 45
<211> 17
<212> DNA
<213> Artificial sequence
<400> 45
ccaggtggaa gaaggta 17
<210> 46
<211> 30
<212> DNA
<213> Artificial sequence
<400> 46
acgttggatg tttccccaag gtcacatagc 30
<210> 47
<211> 30
<212> DNA
<213> Artificial sequence
<400> 47
acgttggatg caatacagca gaacaccgac 30
<210> 48
<211> 16
<212> DNA
<213> Artificial sequence
<400> 48
ccgacggaca taactc 16
<210> 49
<211> 30
<212> DNA
<213> Artificial sequence
<400> 49
acgttggatg tgggtcctgg ggagcagcaa 30
<210> 50
<211> 29
<212> DNA
<213> Artificial sequence
<400> 50
acgttggatg tctctgcact ctgcctctc 29
<210> 51
<211> 15
<212> DNA
<213> Artificial sequence
<400> 51
gagcagcaat gggcg 15
<210> 52
<211> 30
<212> DNA
<213> Artificial sequence
<400> 52
acgttggatg ggtcatgctc aatattcacc 30
<210> 53
<211> 30
<212> DNA
<213> Artificial sequence
<400> 53
acgttggatg aacctgggtc tcccatattg 30
<210> 54
<211> 19
<212> DNA
<213> Artificial sequence
<400> 54
gcccatattg caggcagac 19
<210> 55
<211> 30
<212> DNA
<213> Artificial sequence
<400> 55
acgttggatg gtatccctca gcttcatttg 30
<210> 56
<211> 30
<212> DNA
<213> Artificial sequence
<400> 56
acgttggatg aagcacctgg atggttcttg 30
<210> 57
<211> 22
<212> DNA
<213> Artificial sequence
<400> 57
ttcttctaat tcttaccatc tc 22
<210> 58
<211> 30
<212> DNA
<213> Artificial sequence
<400> 58
acgttggatg aggaagctac aggaggagtc 30
<210> 59
<211> 30
<212> DNA
<213> Artificial sequence
<400> 59
acgttggatg cttagttgta tgcctcagcc 30
<210> 60
<211> 18
<212> DNA
<213> Artificial sequence
<400> 60
ggaggaggag tccacttc 18
<210> 61
<211> 30
<212> DNA
<213> Artificial sequence
<400> 61
acgttggatg gtgtcatgcc cattgtaaag 30
<210> 62
<211> 30
<212> DNA
<213> Artificial sequence
<400> 62
acgttggatg ccacagagaa ggcaggaaaa 30
<210> 63
<211> 23
<212> DNA
<213> Artificial sequence
<400> 63
ggatgattca acagaacttc tga 23
<210> 64
<211> 30
<212> DNA
<213> Artificial sequence
<400> 64
acgttggatg tttgggtgcg ttccctctac 30
<210> 65
<211> 29
<212> DNA
<213> Artificial sequence
<400> 65
acgttggatg ctcacacctg cagcccctg 29
<210> 66
<211> 18
<212> DNA
<213> Artificial sequence
<400> 66
cctgcctccc ctccacac 18
<210> 67
<211> 30
<212> DNA
<213> Artificial sequence
<400> 67
acgttggatg aacatccact gagaaaggtc 30
<210> 68
<211> 30
<212> DNA
<213> Artificial sequence
<400> 68
acgttggatg catcggccaa ctaacatatc 30
<210> 69
<211> 22
<212> DNA
<213> Artificial sequence
<400> 69
attggttatt ttgaaattcc tc 22
<210> 70
<211> 30
<212> DNA
<213> Artificial sequence
<400> 70
acgttggatg atggaaatct ggcccctttg 30
<210> 71
<211> 30
<212> DNA
<213> Artificial sequence
<400> 71
acgttggatg taacattggt ttccgccagc 30
<210> 72
<211> 16
<212> DNA
<213> Artificial sequence
<400> 72
gcccctttgc aaaatt 16
<210> 73
<211> 30
<212> DNA
<213> Artificial sequence
<400> 73
acgttggatg tgactccaca ggtatgtcag 30
<210> 74
<211> 30
<212> DNA
<213> Artificial sequence
<400> 74
acgttggatg ccatggtttc aagtgctctg 30
<210> 75
<211> 16
<212> DNA
<213> Artificial sequence
<400> 75
cgaccctctc ccacac 16
<210> 76
<211> 30
<212> DNA
<213> Artificial sequence
<400> 76
acgttggatg gtgcaaggtg ccatcatttg 30
<210> 77
<211> 30
<212> DNA
<213> Artificial sequence
<400> 77
acgttggatg ttccaggcag acgctttaac 30
<210> 78
<211> 23
<212> DNA
<213> Artificial sequence
<400> 78
cttccgacgc tttaacctct gag 23
<210> 79
<211> 30
<212> DNA
<213> Artificial sequence
<400> 79
acgttggatg acacggagaa ttaggcagag 30
<210> 80
<211> 30
<212> DNA
<213> Artificial sequence
<400> 80
acgttggatg ctgcttttct cttgtgtctg 30
<210> 81
<211> 19
<212> DNA
<213> Artificial sequence
<400> 81
cggcagagaa aagaagcag 19
<210> 82
<211> 30
<212> DNA
<213> Artificial sequence
<400> 82
acgttggatg ttctggcccc tatttgtgtc 30
<210> 83
<211> 30
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<400> 83
acgttggatg ttccctgccc ttgatttaag 30
<210> 84
<211> 21
<212> DNA
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<400> 84
cccattaatc aagacaagac a 21
<210> 85
<211> 30
<212> DNA
<213> Artificial sequence
<400> 85
acgttggatg agatgagaag agactccctg 30
<210> 86
<211> 30
<212> DNA
<213> Artificial sequence
<400> 86
acgttggatg cctgctcttg ggaaaatgtc 30
<210> 87
<211> 22
<212> DNA
<213> Artificial sequence
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tcagtcctta aacaggaatg ta 22
Claims (10)
1. a kind of method based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool, includes the following steps: the genome for detecting sheep to be measured
In nucleotide in the combination of 11 groups of SNP sites at each SNP site, determine that the sheep to be measured corresponds to 11 groups of SNP site groups
11 genotype closed；Then according to determining whether the sheep to be measured is sheep known for its fine thick wool as follows: if the sheep to be measured correspond to it is described
11 genotype of 11 groups of SNP sites combination meet the sheep known for its fine thick wool Reference genotype of respective sets SNP site combination, then described to be measured
Sheep is or candidate is sheep known for its fine thick wool；If 11 genotype that the sheep to be measured corresponds to 11 groups of SNP sites combination meet phase
The non-sheep known for its fine thick wool Reference genotype of SNP site combination should be organized, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
11 groups of SNP site groups are combined into following (a1)-(a11)；(a1) corresponding to 11 groups of SNP site combinations shown in-(a11)
Non- sheep known for its fine thick wool Reference genotype and sheep known for its fine thick wool Reference genotype respectively be (b1)-(b11)；
(a1) contain 5 SNP sites, respectively CM001598.2:69009397, CM001602.2:49927473,
CM001603.2:38707088, CM001603.2:46619514 and CM001606.2:34435705；
(a2) contain 5 SNP sites, respectively CM001582.2:86006851, CM001598.2:69009397,
CM001600.2:45516740, CM001601.2:47737780 and CM001603.2:46619514；
(a3) contain 5 SNP sites, respectively CM001582.2:86006851, CM001583.2:42344562,
CM001584.2:11650210, CM001584.2:12200323 and CM001588.2:9423474；
(a4) contain 5 SNP sites, respectively CM001592.2:57058734, CM001595.2:48968164,
CM001598.2:69009397, CM001601.2:47737780 and CM001603.2:38707088；
(a5) contain 5 SNP sites, respectively CM001585.2:30868868, CM001589.2:75561002,
CM001590.2:56924859, CM001598.2:69009397 and CM001606.2:34435705；
(a6) contain 5 SNP sites, respectively CM001585.2:30868868, CM001588.2:84941340,
CM001598.2:45312379, CM001598.2:69009397 and CM001603.2:46619514；
(a7) contain 5 SNP sites, respectively CM001585.2:30868868, CM001588.2:84941340,
CM001594.2:45477903, CM001594.2:80961940 and CM001603.2:38707088；
(a8) contain 5 SNP sites, respectively CM001585.2:30868868, CM001588.2:84941340,
CM001593.2:1397868, CM001595.2:14087769 and CM001602.2:49927473；
(a9) contain 5 SNP sites, respectively CM001585.2:30868868, CM001588.2:84941340,
CM001592.2:34421625, CM001592.2:53664048 and CM001595.2:48968164；
(a10) contain 5 SNP sites, respectively CM001584.2:79082958, CM001584.2:157466635,
CM001601.2:47737780, CM001602.2:49927473 and CM001603.2:38707088；
(a11) contain 5 SNP sites, respectively CM001584.2:79082958, CM001585.2:5250467,
CM001588.2:84941340, CM001595.2:43390933 and CM001595.2:48968164；
The physical location of each SNP site in 11 groups of SNP sites combination is compared based on sheep known for its fine thick wool full-length genome standard sequence
Determining, the version number of the sheep known for its fine thick wool full-length genome standard sequence is Ovis.Aries V4.0；
CM001582.2:86006851 is located on No. 1 chromosome the 86006851st, and deoxynucleotide is G or T；
CM001583.2:42344562 is located on No. 2 chromosomes the 42344562nd, and deoxynucleotide is T or C；
CM001584.2:11650210 is located on No. 3 chromosomes the 11650210th, and deoxynucleotide is T or C；
CM001584.2:12200323 is located on No. 3 chromosomes the 12200323rd, and deoxynucleotide is C or G；
CM001584.2:79082958 is located on No. 3 chromosomes the 79082958th, and deoxynucleotide is A or G；
CM001584.2:157466635 is located on No. 3 chromosomes the 157466635th, and deoxynucleotide is C or T；
CM001585.2:5250467 is located on No. 4 chromosomes the 5250467th, and deoxynucleotide is C or G；
CM001585.2:30868868 is located on No. 4 chromosomes the 30868868th, and deoxynucleotide is T or C；
CM001588.2:9423474 is located on No. 7 chromosomes the 9423474th, and deoxynucleotide is A or C；
CM001588.2:84941340 is located on No. 7 chromosomes the 84941340th, and deoxynucleotide is A or C；
CM001589.2:75561002 is located on No. 8 chromosomes the 75561002nd, and deoxynucleotide is C or G；
CM001590.2:56924859 is located on No. 9 chromosomes the 56924859th, and deoxynucleotide is A or G；
CM001592.2:34421625 is located on No. 11 chromosomes the 34421625th, and deoxynucleotide is T or C；
CM001592.2:53664048 is located on No. 11 chromosomes the 53664048th, and deoxynucleotide is A or G；
CM001592.2:57058734 is located on No. 11 chromosomes the 57058734th, and deoxynucleotide is C or G；
CM001593.2:1397868 is located on No. 12 chromosomes the 1397868th, and deoxynucleotide is A or G；
CM001594.2:45477903 is located on No. 13 chromosomes the 45477903rd, and deoxynucleotide is A or G；
CM001594.2:80961940 is located on No. 13 chromosomes the 80961940th, and deoxynucleotide is A or C；
CM001595.2:14087769 is located on No. 14 chromosomes the 14087769th, and deoxynucleotide is A or T；
CM001595.2:43390933 is located on No. 14 chromosomes the 43390933rd, and deoxynucleotide is A or G；
CM001595.2:48968164 is located on No. 14 chromosomes the 48968164th, and deoxynucleotide is T or G；
CM001598.2:45312379 is located on No. 17 chromosomes the 45312379th, and deoxynucleotide is T or C；
CM001598.2:69009397 is located on No. 17 chromosomes the 69009397th, and deoxynucleotide is A or C；
CM001600.2:45516740 is located on No. 19 chromosomes the 45516740th, and deoxynucleotide is A or C；
CM001601.2:47737780 is located on No. 20 chromosomes the 47737780th, and deoxynucleotide is A or G；
CM001602.2:49927473 is located on No. 21 chromosomes the 49927473rd, and deoxynucleotide is T or C；
CM001603.2:38707088 is located on No. 22 chromosomes the 38707088th, and deoxynucleotide is T or C；
CM001603.2:46619514 is located on No. 22 chromosomes the 46619514th, and deoxynucleotide is A or G；
CM001606.2:34435705 is located on No. 25 chromosomes the 34435705th, and deoxynucleotide is C or G；
(b1) non-sheep known for its fine thick wool Reference genotype is following any: CC_TT_CC_AA_CG；CC_TC_CC_AA_GG；CC_TT_CC_AA_
GG；CC_TC_TC_AA_GG；AC_TT_CC_AA_GG；AC_TT_CC_AA_CG；CC CC CC AA CG；AC CC TC AA
GG；AC CC CC AA CG；CC CC CC AA GG；CC_TC_TC_AA_CG；CC_TT_TC_AA_GG；CC_TC_CC_AA_
CG；CC_TT_TC_AA_CG；Sheep known for its fine thick wool Reference genotype is following any: CC_CC_TT_AA_CG；AA_CC_CC_AA_CG；CC_
TC_TT_AA_CG；CC_CC_CC_GA_CC；AC_TC_TC_AA_CG；AC_CC_TC_AA_CG；CC_CC_TC_AA_CC；AC_
TC_CC_AA_GG；AA_CC_TC_GA_CG；CC_CC_CC_GG_CG；AA_CC_CC_GG_CC；AC_CC_TC_GA_GG；AA_
CC_TC_AA_CC；CC_CC_TT_GA_GG；AC_CC_CC_AA_CC；AC_CC_TC_GA_CC；
(b2) non-sheep known for its fine thick wool Reference genotype is following any: TT_CC_CC_GG_AA；TT_AC_CC_GG_AA；TT_CC_CC_AG_
AA；TT_AC_AC_AG_AA；TT_CC_AC_AG_AA；TT_CC_AC_GG_AA；TT_CC_CC_AA_AA；Sheep known for its fine thick wool Reference genotype
It is following any: TT_AA_AC_AG_GA；TT_CC_AA_AG_AA；GT_AC_CC_AG_AA；TT_AC_AC_GG_AA；GG_AC_
AC_GG_GA；TT_AC_CC_AG_AA；GT_AC_AC_AG_AA；GG_CC_CC_AG_GG；GG_AC_AC_AA_GA；TT_CC_
AC_GG_GA；GT_AA_AC_AG_AA；GG_CC_AA_AA_GA；GT_AC_AC_AG_GA；TT_AC_AC_AA_GA；TT_CC_
AC_AG_GA；
(b3) non-sheep known for its fine thick wool Reference genotype is following any: TT_TC_TT_CG_AC；TT_TT_TT_GG_AC；TT_TC_TT_GG_
CC；TT_CC_CT_GG_CC；TT_TT_CT_GG_CC；TT_TT_TT_GG_CC；TT_CC_TT_GG_CC；TT_TT_TT_CG_
CC；TT_TC_CT_GG_CC；TT_TC_TT_CG_CC；TT_TC_TT_GG_AC；GT_TT_TT_GG_CC；Sheep known for its fine thick wool Reference genotype
It is following any: TT_CC_CT_CG_CC；TT_TC_CC_GG_CC；TT_TT_CT_CC_CC；GT_CC_CT_CG_AC；GG_CC_
TT_GG_AC；GT_CC_TT_GG_CC；TT_CC_TT_CG_AC；GG_TT_CC_CC_AA；TT_CC_CT_GG_AC；TT_CC_
CC_CG_AA；GT_CC_CT_CG_CC；
(b4) non-sheep known for its fine thick wool Reference genotype is following any: CC_GT_CC_GG_CC；GC_TT_CC_GG_CC；GG_TT_CC_AG_
TC；GG_GT_AC_GG_CC；GC_GT_CC_GG_CC；GC_TT_CC_AG_TC；CC_GT_AC_AG_CC；GC_TT_CC_GG_
TC；GG_TT_AC_GG_TC；GG_TT_CC_GG_CC；CC_TT_CC_AG_CC；CC_TT_AC_GG_CC；GC_TT_CC_AG_
CC；CC_TT_CC_GG_CC；CC_TT_CC_GG_TC；GC_GT_CC_AA_TC；GG_TT_CC_AG_CC；GG_GT_CC_GG_
CC；GC_GT_CC_GG_TC；Sheep known for its fine thick wool Reference genotype is following any: CC_GT_AA_AG_TC；GC_GT_AC_GG_CC；GC_
GT_AC_AG_TC；CC_GT_AA_AA_CC；GC_GG_AA_AG_TC；CC_GT_AC_AG_TT；CC_GG_CC_AA_CC；GC_
GG_AC_GG_TC；CC_TT_AC_GG_TT；CC_GT_AC_AG_TC；CC_GG_CC_AG_TC；CC_GG_AC_AG_TC；CC_
TT_CC_AG_TT；CC_GT_CC_AG_CC；CC_GG_AA_AG_CC；CC_TT_AC_AA_TC；CC_GG_AA_AG_TC；CC_
GT_CC_AA_TT；CC_GG_AC_AG_TT；CC_GT_AC_AA_CC；CC_GT_AC_AA_TC；CC_TT_AC_AG_CC；
(b5) non-sheep known for its fine thick wool Reference genotype is following any: CC GG AA CC CG；CC GG GA AC GG；CC_GG_GA_CC_
GG；CC_CC_GA_CC_CG；CC_CC_AA_CC_GG；TC_GG_GG_AC_GG；CC_CG_GG_CC_CG；CC_CG_GA_CC_
GG；TC_CG_GA_AC_CG；CC_CG_GA_CC_CG；TC_CG_GA_AC_GG；CC_CG_GG_CC_GG；TC_CG_AA_CC_
GG；CC_CG_AA_CC_GG；CC_GG_GG_CC_GG；CC_GG_GA_AC_CG；TC_GG_GG_CC_GG；TC_GG_GG_CC_
CG；TC_GG_GA_CC_CG；TC_CG_GG_CC_CG；TC_GG_GA_CC_GG；Sheep known for its fine thick wool Reference genotype is following any: TC_
CG_GA_AC_CC；TC_CC_GA_CC_CG；TT_CG_AA_AA_CG；CC_CC_AA_CC_CC；TT_CG_GA_AC_CG；TC_
CC_GA_AC_CG；TT_CC_AA_CC_CC；TC_CG_AA_CC_CG；TC_CG_GA_CC_CG；TC_GG_GA_AC_GG；CC_
CC_AA_CC_CG；TT_CG_GA_AA_CC；CC_CC_AA_AA_GG；TT_CC_GA_AC_CG；CC_CC_GA_AC_CC；
(b6) non-sheep known for its fine thick wool Reference genotype is following any: CC_CA_CC_CC_AA；TC_AA_TC_CC_AA；CC_AA_CC_AC_
AA；CC_CA_TC_CC_AA；CC_CA_TT_CC_AA；TC_CA_CC_AC_AA；CC_AA_CC_CC_AA；CC_AA_TC_CC_
AA；TC_AA_CC_CC_AA；TC_CC_TC_CC_AA；CC_AA_TT_CC_AA；CC_CC_TC_CC_AA；TC_CA_CC_CC_
AA；Sheep known for its fine thick wool Reference genotype is following any: TC_CA_TC_AA_GA；TC_AA_TT_CC_AA；TT_CC_TC_AA_AA；TT_
AA_TT_AC_GA；TC_CA_TC_CC_AA；TC_AA_TC_AC_GA；TC_CC_TC_AC_AA；TC_AA_TT_AC_AA；TT_
CC_CC_CC_AA；TC_AA_TC_AC_AA；TC_CA_TT_AC_GA；CC_CA_TT_CC_GA；TT_CC_TT_AA_AA；TC_
CC_TC_CC_GA；TT_CC_TC_AC_GA；CC_CA_TT_AC_AA；
(b7) non-sheep known for its fine thick wool Reference genotype is following any: CC_CA_GG_AA_CC；TC_AA_GG_CA_CC；CC_CA_GG_AA_
TC；TC_CA_GG_AA_CC；CC_AA_GG_CA_CC；TC_CA_GG_CA_CC；TC_AA_GG_CA_TC；CC_AA_GG_CA_
TC；CC_CA_GG_CC_CC；CC_AA_GG_AA_CC；CC_CA_AG_AA_TC；TC_CC_GG_CA_CC；TC_AA_GG_AA_
TC；TC_CA_AG_AA_CC；CC_CC_GG_AA_CC；TC_AA_AG_AA_CC；Sheep known for its fine thick wool Reference genotype is following any: TC_
CA_AG_AA_TC；TC_AA_AG_AA_TC；TT_AA_GG_AA_TC；TT_CC_AG_AA_CC；TT_CC_GG_AA_TC；TT_
AA_AG_AA_TC；TC_CA_AA_AA_TT；TC_CA_AA_AA_CC；TC_CC_AG_AA_TC；TC_CA_GG_AA_TC；TC_
CA_GG_AA_TT；TT_CC_AG_AA_TC；TC_CC_AG_AA_CC；CC_CA_AG_AA_CC；
(b8) non-sheep known for its fine thick wool Reference genotype is following any: CC_CA_GG_AA_TT；CC_AA_GG_AA_TC；CC_CA_AG_AA_
TT；CC_CA_GG_TT_TC；CC_CA_AG_AA_TC；TC_CA_AG_TA_TT；CC_AA_AG_TT_CC；CC_AA_AG_TA_
TC；CC_AA_AA_TA_CC；TC_AA_AG_TT_CC；CC_AA_GG_TT_TC；TC_AA_GG_AA_TT；CC_AA_GG_TT_
CC；CC_AA_AG_TA_CC；TC_AA_AG_TT_TC；CC_CA_AG_TT_CC；CC_AA_AG_TT_TC；CC_AA_AG_AA_
CC；CC_AA_GG_AA_CC；CC_CA_GG_TA_TC；TC_CC_GG_AA_TT；TC_AA_GG_TA_CC；TC_AA_AG_AA_
TC；CC_AA_GG_TT_TT；CC_AA_GG_TA_TT；CC_AA_AA_TA_TC；CC_CA_GG_TT_TT；CC_CC_GG_TA_
CC；TC_AA_GG_TA_TC；TC_CA_GG_TT_TC；CC_AA_GG_TA_TC；TC_AA_AG_AA_TT；Sheep known for its fine thick wool Reference genotype
It is following any: TC_CA_AA_AA_TC；TC_CA_AG_AA_CC；TC_AA_AG_TA_CC；TT_CC_AA_AA_TC；TT_AA_
AA_AA_CC；TC_AA_AG_TA_TC；TC_CC_AG_AA_CC；TC_CA_GG_AA_CC；TC_CA_AG_AA_TC；TC_AA_
AG_AA_CC；TT_CC_GG_AA_CC；TC_CC_AA_AA_CC；CC_CA_AA_AA_TC；TT_CC_AA_AA_CC；TT_AA_
GG_AA_CC；CC_CA_GG_AA_CC；
(b9) non-sheep known for its fine thick wool Reference genotype is following any: CC_CA_TT_GG_GT；TC_AA_TT_GG_GT；CC_AA_TT_AG_
GT；TC_CA_TT_GG_GT；CC_AA_TT_GG_GT；CC_AA_CT_GG_TT；CC_AA_TT_GG_TT；TC_AA_TT_AG_
GT；TC_AA_TT_GG_TT；CC_CA_TT_GG_TT；CC_CA_TT_AG_TT；TC_CC_TT_GG_TT；CC_CC_TT_AG_
GT；TC_CA_TT_AG_TT；Sheep known for its fine thick wool Reference genotype is following any: TC_CA_TT_AG_GT；TT_AA_CT_AG_GT；TT_
CC_TT_AG_GG；TC_AA_TT_AG_TT；TC_AA_TT_GG_GG；TT_CC_CT_AG_GG；TC_CA_TT_GG_TT；TC_
CA_CT_GG_GT；CC_CA_TT_AG_GG；TT_CC_CT_AA_GG；TC_CC_CT_AG_GG；TC_CC_CT_AG_GT；TT_
AA_TT_AA_TT；TT_CC_TT_AA_GT；CC_CA_CT_AG_TT；
(b10) non-sheep known for its fine thick wool Reference genotype is following any: AA_CC_GG_TT_CC；AA_CT_GG_TT_CC；GA_CT_GG_
TT_CC；GA_CC_AG_TC_TC；GA_CC_AG_CC_CC；GA_CT_GG_TC_TC；GA_CC_GG_TC_CC；AA_CT_GG_
CC_CC；GA_CT_AG_CC_CC；AA_CT_AG_CC_CC；GA_TT_GG_CC_CC；AA_CC_AG_CC_CC；AA_CT_AG_
TC_TC；AA_CT_GG_TC_TC；GG_TT_GG_TT_CC；GA_TT_AA_TT_TC；GG_CC_AG_TC_CC；GA_CC_AG_
TT_CC；GA_CT_AG_TC_CC；AA_CC_GG_CC_CC；GA_TT_GG_TC_CC；AA_CC_GG_TC_CC；GA_CT_GG_
TC_CC；AA_CC_GG_TT_TC；Sheep known for its fine thick wool Reference genotype is following any: GA_CT_AG_TC_TC；AA_TT_GG_CC_CC；
GA_CT_AG_CC_TT；GG_TT_AG_TC_TT；GG_TT_GG_TC_TT；GG_CT_GG_TC_CC；GG_CT_AA_CC_CC；
GG_CT_GG_TC_TC；GG_CT_GG_TC_TT；AA_TT_AG_CC_TC；GA_CT_GG_CC_TC；GA_TT_AG_CC_TC；
AA_CT_AA_CC_TC；AA_CT_AG_TC_CC；GG_TT_AG_CC_TC；GG_TT_AG_CC_CC；GG_CT_AG_CC_CC；
GG_CT_AA_CC_TC；GG_TT_GG_TC_TC；GG_TT_AA_CC_CC；GA_TT_AA_CC_TC；GA_TT_AG_CC_CC；
(b11) non-sheep known for its fine thick wool Reference genotype is following any: AA_GC_CA_AG_GT；GA_GC_AA_AG_GT；GG_GC_AA_
GG_GT；AA_CC_CA_AG_TT；GA_GC_CA_AG_TT；AA_CC_CA_GG_GT；GA_GC_AA_GG_GT；GA_CC_AA_
AA_GT；GA_CC_AA_AG_TT；AA_GG_AA_AG_TT；GA_CC_AA_GG_TT；AA_GG_AA_GG_TT；AA_CC_AA_
GG_TT；GA_GC_AA_GG_TT；AA_GC_AA_GG_TT；AA_GC_CA_AG_TT；AA_CC_AA_AG_TT；GA_GC_AA_
AG_TT；AA_GC_AA_AG_TT；AA_CC_CA_GG_TT；GA_CC_CC_AG_TT；AA_CC_AA_AA_TT；GG_CC_AA_
GG_TT；GA_CC_CA_AG_GT；AA_CC_CC_AG_GT；GA_CC_AA_AA_TT；AA_CC_AA_GG_GT；Sheep known for its fine thick wool standard base
Because type is following any: AA_GC_AA_GG_GT；AA_GC_CC_AG_GT；GA_GC_CA_AA_GT；GA_GG_CA_AG_TT；
GA_GG_AA_AG_GG；AA_GG_CC_AG_GG；GG_GC_CC_AA_GT；GG_GC_CA_AG_GT；GG_GC_CC_AA_GG；
GG_GC_CA_AA_TT；GA_GC_CA_AG_GG；GG_GC_CC_AG_GG；GG_CC_CC_AA_GG；GG_GC_CC_AG_GT；
GA_GC_CA_AG_GT；GA_GC_CC_AG_GT；AA_GG_CC_GG_GT.
2. a method of it is following any based on Genotyping identification sheep known for its fine thick wool and non-sheep known for its fine thick wool:
(1) include the following steps: to detect SNP site shown in (a1) described in claim 1 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；Then according to determine as follows the sheep to be measured whether be
Sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b1) described in claim 1, it is described to
Survey sheep is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool mark in (b1) described in claim 1
Quasi- genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(2) include the following steps: to detect SNP site shown in (a2) described in claim 1 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；Then according to determine as follows the sheep to be measured whether be
Sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b2) described in claim 1, it is described to
Survey sheep is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool mark in (b2) described in claim 1
Quasi- genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(3) include the following steps: to detect SNP site shown in (a3) described in claim 1 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；Then according to determine as follows the sheep to be measured whether be
Sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b3) described in claim 1, it is described to
Survey sheep is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool mark in (b3) described in claim 1
Quasi- genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(4) include the following steps: to detect SNP site shown in (a4) described in claim 1 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；Then according to determine as follows the sheep to be measured whether be
Sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b4) described in claim 1, it is described to
Survey sheep is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool mark in (b4) described in claim 1
Quasi- genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(5) include the following steps: to detect SNP site shown in (a5) described in claim 1 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；Then according to determine as follows the sheep to be measured whether be
Sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b5) described in claim 1, it is described to
Survey sheep is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool mark in (b5) described in claim 1
Quasi- genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(6) include the following steps: to detect SNP site shown in (a6) described in claim 1 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；Then according to determine as follows the sheep to be measured whether be
Sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b6) described in claim 1, it is described to
Survey sheep is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool mark in (b6) described in claim 1
Quasi- genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(7) include the following steps: to detect SNP site shown in (a7) described in claim 1 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；Then according to determine as follows the sheep to be measured whether be
Sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b7) described in claim 1, it is described to
Survey sheep is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool mark in (b7) described in claim 1
Quasi- genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(8) include the following steps: to detect SNP site shown in (a8) described in claim 1 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；Then according to determine as follows the sheep to be measured whether be
Sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b8) described in claim 1, it is described to
Survey sheep is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool mark in (b8) described in claim 1
Quasi- genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(9) include the following steps: to detect SNP site shown in (a9) described in claim 1 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；Then according to determine as follows the sheep to be measured whether be
Sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b9) described in claim 1, it is described to
Survey sheep is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-sheep known for its fine thick wool mark in (b9) described in claim 1
Quasi- genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(10) include the following steps: to detect SNP site group shown in (a10) described in claim 1 in the genome of sheep to be measured
Nucleotide in conjunction at each SNP site determines the genotype of the sheep to be measured；Then according to whether determining the sheep to be measured as follows
For sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b10) described in claim 1, institute
State that sheep to be measured is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-beach in (b10) described in claim 1
Sheep Reference genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool；
(11) include the following steps: to detect SNP site group shown in (a11) described in claim 1 in the genome of sheep to be measured
Nucleotide in conjunction at each SNP site determines the genotype of the sheep to be measured；Then according to whether determining the sheep to be measured as follows
For sheep known for its fine thick wool: if the genotype of the sheep to be measured meets the sheep known for its fine thick wool Reference genotype in (b11) described in claim 1, institute
State that sheep to be measured is or candidate is sheep known for its fine thick wool；If the genotype of the sheep to be measured meets the non-beach in (b11) described in claim 1
Sheep Reference genotype, then the sheep to be measured is or candidate is non-sheep known for its fine thick wool.
3. a kind of method based on Genotyping identification sheep known for its fine thick wool, includes the following steps: to detect 11 groups of SNP in the genome of sheep to be measured
Nucleotide in Sites Combination at each SNP site determines that the sheep to be measured corresponds to 11 bases of 11 groups of SNP sites combination
Because of type；If 11 genotype that the sheep to be measured corresponds to 11 groups of SNP sites combination meet respective sets SNP site group
The sheep known for its fine thick wool Reference genotype of conjunction, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
11 groups of SNP site groups are combined into (a1)-(a11) described in claim 1；(a1) 11 groups of SNP shown in-(a11)
Sheep known for its fine thick wool Reference genotype corresponding to Sites Combination respectively is the sheep known for its fine thick wool mark in (b1)-(b11) described in claim 1
Quasi- genotype.
4. a kind of method based on Genotyping identification sheep known for its fine thick wool is following any:
(1) include the following steps: to detect SNP site shown in (a1) described in claim 3 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right and wants
The sheep known for its fine thick wool Reference genotype in (b1) described in 3 is sought, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(2) include the following steps: to detect SNP site shown in (a2) described in claim 3 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right and wants
The sheep known for its fine thick wool Reference genotype in (b2) described in 3 is sought, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(3) include the following steps: to detect SNP site shown in (a3) described in claim 3 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right and wants
The sheep known for its fine thick wool Reference genotype in (b3) described in 3 is sought, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(4) include the following steps: to detect SNP site shown in (a4) described in claim 3 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right and wants
The sheep known for its fine thick wool Reference genotype in (b4) described in 3 is sought, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(5) include the following steps: to detect SNP site shown in (a5) described in claim 3 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right and wants
The sheep known for its fine thick wool Reference genotype in (b5) described in 3 is sought, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(6) include the following steps: to detect SNP site shown in (a6) described in claim 3 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right and wants
The sheep known for its fine thick wool Reference genotype in (b6) described in 3 is sought, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(7) include the following steps: to detect SNP site shown in (a7) described in claim 3 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right and wants
The sheep known for its fine thick wool Reference genotype in (b7) described in 3 is sought, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(8) include the following steps: to detect SNP site shown in (a8) described in claim 3 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right and wants
The sheep known for its fine thick wool Reference genotype in (b8) described in 3 is sought, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(9) include the following steps: to detect SNP site shown in (a9) described in claim 3 in the genome of sheep to be measured to combine
In nucleotide at each SNP site, determine the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right and wants
The sheep known for its fine thick wool Reference genotype in (b9) described in 3 is sought, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(10) include the following steps: to detect SNP site group shown in (a10) described in claim 3 in the genome of sheep to be measured
Nucleotide in conjunction at each SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right
It is required that the sheep known for its fine thick wool Reference genotype in (b10) described in 3, then the sheep to be measured is or candidate is sheep known for its fine thick wool；
(11) include the following steps: to detect SNP site group shown in (a11) described in claim 3 in the genome of sheep to be measured
Nucleotide in conjunction at each SNP site determines the genotype of the sheep to be measured；If the genotype of the sheep to be measured meets right
It is required that the sheep known for its fine thick wool Reference genotype in (b11) described in 3, then the sheep to be measured is or candidate is sheep known for its fine thick wool.
5. applying (A1) or (A2) as follows:
(A1) for detecting the nucleosides in sheep genome in 11 groups of SNP site combinations described in claim 1 at each SNP site
Application of the substance of acid in identification sheep known for its fine thick wool and/or non-sheep known for its fine thick wool, or preparing the reagent for identifying sheep known for its fine thick wool and/or non-sheep known for its fine thick wool
Application in box；
(A2) each SNP of any one group in 11 groups of SNP sites combinations described in claim 1 are used to detect in sheep genome
The identification sheep known for its fine thick wool of nucleotide at point and/or the application in non-sheep known for its fine thick wool, or in preparation for identifying sheep known for its fine thick wool and/or non-beach
Application in the kit of sheep.
6. application according to claim 5, it is characterised in that: the substance is for designed by SNP site to be detected
PCR primer and/or sequencing primer.
7. application according to claim 6, it is characterised in that:
Primer for detecting the CM001582.2:86006851 is two lists shown in SEQ ID No.1 and SEQ ID No.2
Chain DNA；
Primer for detecting the CM001583.2:42344562 is two lists shown in SEQ ID No.4 and SEQ ID No.5
Chain DNA；
Primer for detecting the CM001584.2:11650210 is two lists shown in SEQ ID No.7 and SEQ ID No.8
Chain DNA；
Primer for detecting the CM001584.2:12200323 is two shown in SEQ ID No.10 and SEQ ID No.11
Single stranded DNA；
Primer for detecting the CM001584.2:79082958 is two shown in SEQ ID No.13 and SEQ ID No.14
Single stranded DNA；
Primer for detecting the CM001584.2:157466635 is two shown in SEQ ID No.16 and SEQ ID No.17
Single stranded DNA；
Primer for detecting the CM001585.2:5250467 is two shown in SEQ ID No.19 and SEQ ID No.20
Single stranded DNA；
Primer for detecting the CM001585.2:30868868 is two shown in SEQ ID No.22 and SEQ ID No.23
Single stranded DNA；
Primer for detecting the CM001588.2:9423474 is two shown in SEQ ID No.25 and SEQ ID No.26
Single stranded DNA；
Primer for detecting the CM001588.2:84941340 is two shown in SEQ ID No.28 and SEQ ID No.29
Single stranded DNA；
Primer for detecting the CM001589.2:75561002 is two shown in SEQ ID No.31 and SEQ ID No.32
Single stranded DNA；
Primer for detecting the CM001590.2:56924859 is two shown in SEQ ID No.34 and SEQ ID No.35
Single stranded DNA；
Primer for detecting the CM001592.2:34421625 is two shown in SEQ ID No.37 and SEQ ID No.38
Single stranded DNA；
Primer for detecting the CM001592.2:53664048 is two shown in SEQ ID No.40 and SEQ ID No.41
Single stranded DNA；
Primer for detecting the CM001592.2:57058734 is two shown in SEQ ID No.43 and SEQ ID No.44
Single stranded DNA；
Primer for detecting the CM001593.2:1397868 is two shown in SEQ ID No.46 and SEQ ID No.47
Single stranded DNA；
Primer for detecting the CM001594.2:45477903 is two shown in SEQ ID No.49 and SEQ ID No.50
Single stranded DNA；
Primer for detecting the CM001594.2:80961940 is two shown in SEQ ID No.52 and SEQ ID No.53
Single stranded DNA；
Primer for detecting the CM001595.2:14087769 is two shown in SEQ ID No.55 and SEQ ID No.56
Single stranded DNA；
Primer for detecting the CM001595.2:43390933 is two shown in SEQ ID No.58 and SEQ ID No.59
Single stranded DNA；
Primer for detecting the CM001595.2:48968164 is two shown in SEQ ID No.61 and SEQ ID No.62
Single stranded DNA；
Primer for detecting the CM001598.2:45312379 is two shown in SEQ ID No.64 and SEQ ID No.65
Single stranded DNA；
Primer for detecting the CM001598.2:69009397 is two shown in SEQ ID No.67 and SEQ ID No.68
Single stranded DNA；
Primer for detecting the CM001600.2:45516740 is two shown in SEQ ID No.70 and SEQ ID No.71
Single stranded DNA；
Primer for detecting the CM001601.2:47737780 is two shown in SEQ ID No.73 and SEQ ID No.74
Single stranded DNA；
Primer for detecting the CM001602.2:49927473 is two shown in SEQ ID No.76 and SEQ ID No.77
Single stranded DNA；
Primer for detecting the CM001603.2:38707088 is two shown in SEQ ID No.79 and SEQ ID No.80
Single stranded DNA；
Primer for detecting the CM001603.2:46619514 is two shown in SEQ ID No.82 and SEQ ID No.83
Single stranded DNA；
Primer for detecting the CM001606.2:34435705 is two shown in SEQ ID No.85 and SEQ ID No.86
Single stranded DNA.
8. application according to claim 7, it is characterised in that:
Primer for detecting the CM001582.2:86006851 further includes single stranded DNA shown in SEQ ID No.3；
Primer for detecting the CM001583.2:42344562 further includes single stranded DNA shown in SEQ ID No.6；
Primer for detecting the CM001584.2:11650210 further includes single stranded DNA shown in SEQ ID No.9；
Primer for detecting the CM001584.2:12200323 further includes single stranded DNA shown in SEQ ID No.12；
Primer for detecting the CM001584.2:79082958 further includes single stranded DNA shown in SEQ ID No.15；
Primer for detecting the CM001584.2:157466635 further includes single stranded DNA shown in SEQ ID No.18；
Primer for detecting the CM001585.2:5250467 further includes single stranded DNA shown in SEQ ID No.21；
Primer for detecting the CM001585.2:30868868 further includes single stranded DNA shown in SEQ ID No.24；
Primer for detecting the CM001588.2:9423474 further includes single stranded DNA shown in SEQ ID No.27；
Primer for detecting the CM001588.2:84941340 further includes single stranded DNA shown in SEQ ID No.30；
Primer for detecting the CM001589.2:75561002 further includes single stranded DNA shown in SEQ ID No.33；
Primer for detecting the CM001590.2:56924859 further includes single stranded DNA shown in SEQ ID No.36；
Primer for detecting the CM001592.2:34421625 further includes single stranded DNA shown in SEQ ID No.39；
Primer for detecting the CM001592.2:53664048 further includes single stranded DNA shown in SEQ ID No.42；
Primer for detecting the CM001592.2:57058734 further includes single stranded DNA shown in SEQ ID No.45；
Primer for detecting the CM001593.2:1397868 further includes single stranded DNA shown in SEQ ID No.48；
Primer for detecting the CM001594.2:45477903 further includes single stranded DNA shown in SEQ ID No.51；
Primer for detecting the CM001594.2:80961940 further includes single stranded DNA shown in SEQ ID No.54；
Primer for detecting the CM001595.2:14087769 further includes single stranded DNA shown in SEQ ID No.57；
Primer for detecting the CM001595.2:43390933 further includes single stranded DNA shown in SEQ ID No.60；
Primer for detecting the CM001595.2:48968164 further includes single stranded DNA shown in SEQ ID No.63；
Primer for detecting the CM001598.2:45312379 further includes single stranded DNA shown in SEQ ID No.66；
Primer for detecting the CM001598.2:69009397 further includes single stranded DNA shown in SEQ ID No.69；
Primer for detecting the CM001600.2:45516740 further includes single stranded DNA shown in SEQ ID No.72；
Primer for detecting the CM001601.2:47737780 further includes single stranded DNA shown in SEQ ID No.75；
Primer for detecting the CM001602.2:49927473 further includes single stranded DNA shown in SEQ ID No.78；
Primer for detecting the CM001603.2:38707088 further includes single stranded DNA shown in SEQ ID No.81；
Primer for detecting the CM001603.2:46619514 further includes single stranded DNA shown in SEQ ID No.84；
Primer for detecting the CM001606.2:34435705 further includes single stranded DNA shown in SEQ ID No.87.
9. primer set, for as follows (B1) or (B2):
(B1) for detecting the nucleosides in sheep genome in 11 groups of SNP site combinations described in claim 1 at each SNP site
The primer set of acid；
(B2) each SNP of any one group in 11 groups of SNP sites combinations described in claim 1 are used to detect in sheep genome
The primer set of nucleotide at point；
For detecting the primer of each SNP site as described in claim 7 or 8.
10. applying (C1) or (C2) as follows:
(C1) nucleotide in sheep genome in 11 groups of SNP site combinations described in claim 1 at each SNP site is polymorphic
Property identification sheep known for its fine thick wool and/or non-sheep known for its fine thick wool in application；
(C2) core in sheep genome in 11 groups of SNP site combinations described in claim 1 at any one group of each SNP site
Application of the polymorphism of thuja acid in identification sheep known for its fine thick wool and/or non-sheep known for its fine thick wool.
Priority Applications (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810409670.7A CN110438234B (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214561.0A CN116804222A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep and non-sheep based on SNP molecular markers |
| CN202310214552.1A CN116144793A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214555.5A CN116219038A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214557.4A CN116479137A (en) | 2018-05-02 | 2018-05-02 | Method for identifying Tan sheep based on SNP locus combination |
| CN202310214560.6A CN116287306A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on basis of genotyping |
| CN202310214547.0A CN116219037A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201810409670.7A CN110438234B (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202310214557.4A Division CN116479137A (en) | 2018-05-02 | 2018-05-02 | Method for identifying Tan sheep based on SNP locus combination |
| CN202310214552.1A Division CN116144793A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214555.5A Division CN116219038A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214561.0A Division CN116804222A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep and non-sheep based on SNP molecular markers |
| CN202310214547.0A Division CN116219037A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214560.6A Division CN116287306A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on basis of genotyping |
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| CN202310214552.1A Pending CN116144793A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214561.0A Pending CN116804222A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep and non-sheep based on SNP molecular markers |
| CN202310214560.6A Pending CN116287306A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on basis of genotyping |
| CN202310214547.0A Pending CN116219037A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214555.5A Pending CN116219038A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214557.4A Pending CN116479137A (en) | 2018-05-02 | 2018-05-02 | Method for identifying Tan sheep based on SNP locus combination |
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| CN202310214561.0A Pending CN116804222A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep and non-sheep based on SNP molecular markers |
| CN202310214560.6A Pending CN116287306A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on basis of genotyping |
| CN202310214547.0A Pending CN116219037A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214555.5A Pending CN116219038A (en) | 2018-05-02 | 2018-05-02 | Method for identifying sheep on beach and sheep on non-beach based on genotyping |
| CN202310214557.4A Pending CN116479137A (en) | 2018-05-02 | 2018-05-02 | Method for identifying Tan sheep based on SNP locus combination |
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| CN116479137A (en) | 2023-07-25 |
| CN116144793A (en) | 2023-05-23 |
| CN116287306A (en) | 2023-06-23 |
| CN110438234B (en) | 2023-05-26 |
| CN116804222A (en) | 2023-09-26 |
| CN116219038A (en) | 2023-06-06 |
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