CN110438234B - Method for identifying sheep on beach and sheep on non-beach based on genotyping - Google Patents

Abstract

The invention discloses a method for identifying sheep and non-sheep based on genotyping. The method comprises the following steps: detecting the nucleotide of each SNP locus in 11 groups of SNP locus combinations in the genome of the sheep to be detected, and determining 11 genotypes of the sheep to be detected corresponding to the 11 groups of SNP locus combinations; and then determining whether the sheep to be tested is a Tan sheep according to the following steps: if the 11 genotypes of the 11 SNP locus combinations correspond to the standard genotypes of the Tan sheep of the corresponding SNP locus combinations, the sheep to be detected is or is candidate to be Tan sheep; and if the 11 genotypes of the 11 SNP locus combinations correspond to the standard genotypes of the non-Tan sheep of the corresponding SNP locus combinations, the sheep to be tested is or is candidate to be the non-Tan sheep. The invention successfully obtains SNP locus combination for identifying the Tan sheep and the non-Tan sheep, and provides support for the identification, the seed conservation and the genetic breeding of the Tan sheep in the future.

Description

A method for identifying Tan sheep and non-Tan sheep based on genotyping

Technical Field

The invention relates to the field of biotechnology, and in particular to a method for identifying Tan sheep and non-Tan sheep based on genotyping.

Background Art

Tan sheep is an important livestock resource in Ningxia, with an annual output of 6 million heads. It is highly respected by the market for its unique flavor and meat quality and is an important brand in Ningxia. However, due to the recent emergence of hybrids of Tan sheep and other sheep breeds being sold as Tan sheep, the Tan sheep brand has been greatly damaged. In order to stabilize the quality of Tan sheep, rebuild the brand image of Tan sheep, and build Tan sheep into a nationally famous brand, we have identified Tan sheep and non-Tan sheep to lay the foundation for the identification and conservation of Tan sheep and genetic breeding in the future.

In recent years, with the development of molecular biology, gene detection technology has also made great progress in biological species identification. Gene detection technology is widely used in animal and plant species identification because it can identify almost all biological materials. Gene detection technology for identifying animal and plant species is the fastest-growing and most accurate technical means in species identification.

Whole genome resequencing is the whole genome sequencing of individuals with known genome sequences. With the decrease in sequencing costs and the increase in species with known genome sequences, resequencing has become one of the most effective methods in animal and plant breeding research. Sequenom SNP typing technology is based on the principle of primer extension reaction, combining sensitive PCR amplification and reliable MALDI-TOF mass spectrometry technology to achieve genotyping detection. This technology is particularly suitable for verifying the results of whole genome research.

Summary of the invention

The purpose of the present invention is to provide a method for identifying Tan sheep and non-Tan sheep based on genotyping.

In a first aspect, the present invention claims a method for identifying Tan sheep and non-Tan sheep based on genotyping.

The method for identifying Tan sheep and non-Tan sheep based on genotyping provided by the present invention may be any of the following:

The first method is a method for identification based on 11 groups of SNP site combinations. The method may specifically include the following steps: detecting the nucleotides at each SNP site in the 11 groups of SNP site combinations in the genome of the sheep to be tested, and determining the 11 genotypes of the sheep to be tested corresponding to the 11 groups of SNP site combinations; then determining whether the sheep to be tested is a Tan sheep as follows: if the 11 genotypes of the sheep to be tested corresponding to the 11 groups of SNP site combinations all meet the Tan sheep standard genotypes of the corresponding groups of SNP site combinations, then the sheep to be tested is or is a candidate for being a Tan sheep; if the 11 genotypes of the sheep to be tested corresponding to the 11 groups of SNP site combinations all meet the non-Tan sheep standard genotypes of the corresponding groups of SNP site combinations, then the sheep to be tested is or is a candidate for being a non-Tan sheep.

The 11 groups of SNP site combinations are as follows (a1)-(a11); the non-tan sheep standard genotype and the tan sheep standard genotype corresponding to the 11 groups of SNP site combinations shown in (a1)-(a11) are respectively (b1)-(b11). That is, the non-tan sheep standard genotype and the tan sheep standard genotype corresponding to the SNP site combination shown in (a1) are (b1); the non-tan sheep standard genotype and the tan sheep standard genotype corresponding to the SNP site combination shown in (a2) are (b2); ... the non-tan sheep standard genotype and the tan sheep standard genotype corresponding to the SNP site combination shown in (a11) are (b11).

(a1) contains 5 SNP sites, namely CM001598.2:69009397, CM001602.2:49927473, CM001603.2:38707088, CM001603.2:46619514 and CM001606.2:34435705;

(a2) contains 5 SNP sites, namely CM001582.2:86006851, CM001598.2:69009397, CM001600.2:45516740, CM001601.2:47737780 and CM001603.2:46619514;

(a3) contains 5 SNP sites, namely CM001582.2:86006851, CM001583.2:42344562, CM001584.2:11650210, CM001584.2:12200323 and CM001588.2:9423474;

(a4) contains 5 SNP sites, namely CM001592.2:57058734, CM001595.2:48968164, CM001598.2:69009397, CM001601.2:47737780 and CM001603.2:38707088;

(a5) contains 5 SNP sites, namely CM001585.2:30868868, CM001589.2:75561002, CM001590.2:56924859, CM001598.2:69009397 and CM001606.2:34435705;

(a6) contains 5 SNP sites, namely CM001585.2:30868868, CM001588.2:84941340, CM001598.2:45312379, CM001598.2:69009397 and CM001603.2:46619514;

(a7) contains 5 SNP sites, namely CM001585.2:30868868, CM001588.2:84941340, CM001594.2:45477903, CM001594.2:80961940 and CM001603.2:38707088;

(a8) contains 5 SNP sites, namely CM001585.2:30868868, CM001588.2:84941340, CM001593.2:1397868, CM001595.2:14087769 and CM001602.2:49927473;

(a9) contains 5 SNP sites, namely CM001585.2:30868868, CM001588.2:84941340, CM001592.2:34421625, CM001592.2:53664048 and CM001595.2:48968164;

(a10) contains 5 SNP sites, namely CM001584.2:79082958, CM001584.2:157466635, CM001601.2:47737780, CM001602.2:49927473 and CM001603.2:38707088;

(a11) contains 5 SNP sites, namely CM001584.2:79082958, CM001585.2:5250467, CM001588.2:84941340, CM001595.2:43390933 and CM001595.2:48968164;

The physical position of each SNP site in the 11 groups of SNP site combinations is determined based on the comparison of the Tan sheep whole genome standard sequence, the version number of the Tan sheep whole genome standard sequence is Ovis.Aries V4.0; there are 29 SNP sites in total, as follows:

CM001582.2:86006851 is located at position 86006851 on chromosome 1, and its deoxynucleotide is G or T;

CM001583.2:42344562 is located at position 42344562 on chromosome 2, and its deoxynucleotide is T or C;

CM001584.2:11650210 is located at position 11650210 on chromosome 3, and its deoxynucleotide is T or C;

CM001584.2:12200323 is located at position 12200323 on chromosome 3, and its deoxynucleotide is C or G;

CM001584.2:79082958 is located at position 79082958 on chromosome 3, and its deoxynucleotide is A or G;

CM001584.2:157466635 is located at position 157466635 on chromosome 3, and its deoxynucleotide is C or T;

CM001585.2:5250467 is located at position 5250467 on chromosome 4, and its deoxynucleotide is C or G;

CM001585.2:30868868 is located at position 30868868 on chromosome 4, and its deoxynucleotide is T or C;

CM001588.2:9423474 is located at position 9423474 on chromosome 7, and its deoxynucleotide is A or C;

CM001588.2:84941340 is located at position 84941340 on chromosome 7, and its deoxynucleotide is A or C;

CM001589.2:75561002 is located at position 75561002 on chromosome 8, and its deoxynucleotide is C or G;

CM001590.2:56924859 is located at position 56924859 on chromosome 9, and its deoxynucleotide is A or G;

CM001592.2:34421625 is located at position 34421625 on chromosome 11, and its deoxynucleotides are T or C;

CM001592.2:53664048 is located at position 53664048 on chromosome 11, and its deoxynucleotide is A or G;

CM001592.2:57058734 is located at position 57058734 on chromosome 11, and its deoxynucleotide is C or G;

CM001593.2:1397868 is located at position 1397868 on chromosome 12, and its deoxynucleotide is A or G;

CM001594.2:45477903 is located at position 45477903 on chromosome 13, and its deoxynucleotide is A or G;

CM001594.2:80961940 is located at position 80961940 on chromosome 13, and its deoxynucleotide is A or C;

CM001595.2:14087769 is located at position 14087769 on chromosome 14, and its deoxynucleotides are A or T;

CM001595.2:43390933 is located at position 43390933 on chromosome 14, and its deoxynucleotide is A or G;

CM001595.2:48968164 is located at position 48968164 on chromosome 14, and its deoxynucleotide is T or G;

CM001598.2:45312379 is located at position 45312379 on chromosome 17, and its deoxynucleotides are T or C;

CM001598.2:69009397 is located at position 69009397 on chromosome 17, and its deoxynucleotide is A or C;

CM001600.2:45516740 is located at position 45516740 on chromosome 19, and its deoxynucleotide is A or C;

CM001601.2:47737780 is located at position 47737780 on chromosome 20, and its deoxynucleotide is A or G;

CM001602.2:49927473 is located at position 49927473 on chromosome 21, and its deoxynucleotides are T or C;

CM001603.2:38707088 is located at position 38707088 on chromosome 22, and its deoxynucleotides are T or C;

CM001603.2:46619514 is located at position 46619514 on chromosome 22, and its deoxynucleotide is A or G;

CM001606.2:34435705 is located at position 34435705 on chromosome 25, and its deoxynucleotide is C or G;

Of course, the version number of the Tan sheep whole genome standard sequence used in the actual operation process is not limited to Ovis.Aries V4.0. No matter which version number of the Tan sheep whole genome standard sequence is used, as long as the SNP sites involved are the same as the 29 SNP sites mentioned above in the present invention, they all fall within the protection scope of the present invention.

(b1) Non-tan sheep standard genotype is any of the following: 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; Tan sheep standard genotype is any of the following : 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_AA_CC; CC_CC_TT_GA_GG; AC_CC_CC_AA_CC; AC_CC_TC_GA_CC.

(b2) The non-standard genotype of Tan sheep is any of the following: 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; The standard genotype of Tan sheep is any of the following: 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) The non-tan sheep standard genotype is any of the following: 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; the standard genotype of Tan sheep is any of the following: 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) The non-standard genotype of Tan sheep is any of the following: 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; The standard genotype of Tan sheep is any of the following: CC_GT_A A_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; 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) The standard genotype of non-tan sheep is any of the following: 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; _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; The standard genotype of Tan sheep is any of the following: 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) The non-standard genotype of Tan sheep is any of the following: 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; The standard genotype of Tan sheep is any of the following: TC_CA_TC C_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) The non-tan sheep standard genotype is any of the following: 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; the standard genotype of Tan sheep is any of the following: 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) The non-tan sheep standard genotype is any of the following: 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_A A_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; 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; Tan sheep standard genotype is any of the following: TC_CA_AA_AA_TC; TC_CA_AG_AA_CC; TC_AA_AG_TA_CC; TT_CC_A A_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-Tan sheep standard genotype is any of the following: 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; Tan sheep standard genotype The cause type is any of the following: 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) The non-tan sheep standard genotype is any of the following: 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; CC; AA_CC_GG_TT_TC; Tan sheep standard genotype is any of the following: 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) The standard genotype of non-tan sheep is any of the following: 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; Tan sheep standard genotype is any of the following: 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_C C_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.

The genotypes _shown in ( _b1 ) to ₍ b11) above are expressed as _{X1X2_X3X4_X5X6_X7X8_X9X10} , _{wherein X1} to _X10 all represent A or C or _T or _G , _{X1X2 represents} the nucleotide at the first SNP site in the SNP site combination (e.g., AA _represents the homozygous nucleotide at the SNP site; e.g., AT represents the _{heterozygous nucleotide at the SNP site), X3X4 represents the} nucleotide at the second SNP site in the SNP site combination, ..., _X9X10 represents the nucleotide at the fifth SNP site in _the SNP site combination _.

The second method is based on a combination of SNP sites for identification. This method can be any of the following:

(1) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a1) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b1) above, the sheep to be tested is or is a candidate for being a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b1) above, the sheep to be tested is or is a candidate for being a non-Tan sheep.

(2) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a2) in the genome of the sheep to be tested to determine the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b2), the sheep to be tested is or is a candidate for being a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b2), the sheep to be tested is or is a candidate for being a non-Tan sheep.

(3) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a3) in the genome of the sheep to be tested to determine the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b3), the sheep to be tested is or is a candidate for being a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b3), the sheep to be tested is or is a candidate for being a non-Tan sheep.

(4) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a4) in the genome of the sheep to be tested to determine the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b4), the sheep to be tested is or is a candidate for being a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b4) described above, the sheep to be tested is or is a candidate for being a non-Tan sheep.

(5) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a5) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b5) above, the sheep to be tested is or is a candidate for being a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b5) above, the sheep to be tested is or is a candidate for being a non-Tan sheep.

(6) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a6) above in the genome of the sheep to be tested to determine the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b6) above, the sheep to be tested is or is a candidate to be a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b6) above, the sheep to be tested is or is a candidate to be a non-Tan sheep.

(7) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a7) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b7) above, the sheep to be tested is or is a candidate to be a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b7) above, the sheep to be tested is or is a candidate to be a non-Tan sheep.

(8) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a8) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b8) above, the sheep to be tested is or is a candidate to be a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b8) above, the sheep to be tested is or is a candidate to be a non-Tan sheep.

(9) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a9) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b9) above, the sheep to be tested is or is a candidate to be a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b9) above, the sheep to be tested is or is a candidate to be a non-Tan sheep.

(10) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a10) mentioned above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b10) mentioned above, the sheep to be tested is or is a candidate to be a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b10) mentioned above, the sheep to be tested is or is a candidate to be a non-Tan sheep.

(11) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a11) mentioned above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; determining whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested conforms to the Tan sheep standard genotype in (b11) mentioned above, the sheep to be tested is or is a candidate to be a Tan sheep; if the genotype of the sheep to be tested conforms to the non-Tan sheep standard genotype in (b11) mentioned above, the sheep to be tested is or is a candidate to be a non-Tan sheep.

In a second aspect, the present invention claims a method for identifying Tan sheep based on genotyping.

The method for identifying Tan sheep based on genotyping provided by the present invention may be any of the following:

The first method is a method for identification based on 11 groups of SNP site combinations. The method may include the following steps: detecting the nucleotides at each SNP site in the 11 groups of SNP site combinations in the genome of the sheep to be tested, and determining the 11 genotypes of the sheep to be tested corresponding to the 11 groups of SNP site combinations; if the 11 genotypes of the sheep to be tested corresponding to the 11 groups of SNP site combinations all meet the standard genotypes of the corresponding groups of SNP site combinations, then the sheep to be tested is or is a candidate for being a Tan sheep.

Among them, the 11 groups of SNP site combinations are (a1)-(a11) mentioned above; the standard genotypes of Tan sheep corresponding to the 11 groups of SNP site combinations shown in (a1)-(a11) are respectively the standard genotypes of Tan sheep in (b1)-(b11) mentioned above.

The second method is based on a combination of SNP sites for identification. This method can be any of the following:

(1) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a1) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of Tan sheep in (b1) above, then the sheep to be tested is or is a candidate for being Tan sheep.

(2) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a2) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b2) above, then the sheep to be tested is or is a candidate for being a Tan sheep.

(3) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a3) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b3) above, then the sheep to be tested is or is a candidate for being a Tan sheep.

(4) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a4) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b4) above, then the sheep to be tested is or is a candidate for being a Tan sheep.

(5) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a5) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b5) above, then the sheep to be tested is or is a candidate for being a Tan sheep.

(6) comprising the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a6) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b6) above, then the sheep to be tested is or is a candidate for being a Tan sheep.

(7) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a7) above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b7) above, then the sheep to be tested is or is a candidate for being a Tan sheep.

(8) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a8) mentioned above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b8) mentioned above, then the sheep to be tested is or is a candidate to be a Tan sheep.

(9) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a9) mentioned above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b9) mentioned above, then the sheep to be tested is or is a candidate for being a Tan sheep.

(10) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a10) mentioned above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b10) mentioned above, then the sheep to be tested is or is a candidate for being a Tan sheep.

(11) comprises the following steps: detecting the nucleotides at each SNP site in the SNP site combination shown in (a11) mentioned above in the genome of the sheep to be tested, and determining the genotype of the sheep to be tested; if the genotype of the sheep to be tested conforms to the standard genotype of the Tan sheep in (b11) mentioned above, then the sheep to be tested is or is a candidate to be a Tan sheep.

In the methods described in the first aspect and the second aspect, the sheep to be tested may specifically be Tan sheep or non-Tan sheep (such as Mongolian sheep).

In the methods of the first and second aspects, the specific method for detecting the nucleotide at the SNP site in the genome of the sheep to be tested can be a whole genome resequencing method. Further, the DNA sample used for the whole genome resequencing method is taken from the blood or ear tissue of the sheep to be tested.

In a third aspect, the present invention also claims protection for the following applications.

The first one is the use of a substance for detecting nucleotides at each SNP site in the 11 groups of SNP site combinations mentioned above in the sheep genome (or a substance for detecting nucleotides at the 29 SNP sites mentioned above in the sheep genome) in identifying Tan sheep and/or non-Tan sheep, or in the preparation of a kit for identifying Tan sheep and/or non-Tan sheep.

The second type is the use of a material for detecting nucleotides at each SNP site in any of the 11 groups of SNP site combinations mentioned above in the sheep genome for identifying Tan sheep and/or non-Tan sheep, or for preparing a kit for identifying Tan sheep and/or non-Tan sheep.

In the above two applications, the substance may specifically be a PCR primer and/or a sequencing primer designed for the SNP site to be detected.

Further, the primers used to detect the CM001582.2:86006851 are two single-stranded DNAs shown in SEQ ID No.1 and SEQ ID No.2; the primers used to detect the CM001583.2:42344562 are two single-stranded DNAs shown in SEQ ID No.4 and SEQ ID No.5; the primers used to detect the CM001584.2:11650210 are two single-stranded DNAs shown in SEQ ID No.7 and SEQ ID No.8; the primers used to detect the CM001584.2:12200323 are two single-stranded DNAs shown in SEQ ID No.10 and SEQ ID No.11; the primers used to detect the CM001584.2:79082958 are two single-stranded DNAs shown in SEQ ID No.13 and SEQ ID No.14; the primers used to detect the CM001584.2:157466635 are SEQ ID No.16 and SEQ ID No.17; the primers used to detect the CM001585.2:5250467 are the two single-stranded DNAs shown in SEQ ID No.19 and SEQ ID No.20; the primers used to detect the CM001585.2:30868868 are the two single-stranded DNAs shown in SEQ ID No.22 and SEQ ID No.23; the primers used to detect the CM001588.2:9423474 are the two single-stranded DNAs shown in SEQ ID No.25 and SEQ ID No.26; the primers used to detect the CM001588.2:84941340 are the two single-stranded DNAs shown in SEQ ID No.28 and SEQ ID No.29; the primers used to detect the CM001589.2:75561002 are the two single-stranded DNAs shown in SEQ ID No.31 and SEQ ID The primers used to detect CM001590.2:56924859 are the two single-stranded DNAs shown in SEQ ID No.34 and SEQ ID No.35; the primers used to detect CM001592.2:34421625 are the two single-stranded DNAs shown in SEQ ID No.37 and SEQ ID No.38; the primers used to detect CM001592.2:53664048 are the two single-stranded DNAs shown in SEQ ID No.40 and SEQ ID No.41; the primers used to detect CM001592.2:57058734 are the two single-stranded DNAs shown in SEQ ID No.43 and SEQ ID No.44; the primers used to detect CM001593.2:1397868 are the two single-stranded DNAs shown in SEQ ID No.46 and SEQ ID No. The primers used to detect the CM001594.2:45477903 are the two single-stranded DNAs shown in SEQ ID No.49 and SEQ ID No.50; the primers used to detect the CM001594.2:80961940 are the two single-stranded DNAs shown in SEQ ID No.52 and SEQ ID No.53; the primers used to detect the CM001595.2:14087769 are the two single-stranded DNAs shown in SEQ ID No.55 and SEQ ID No.56; the primers used to detect the CM001595.2:43390933 are the two single-stranded DNAs shown in SEQ ID No.58 and SEQ ID No.59; the primers used to detect the CM001595.2:48968164 are the two single-stranded DNAs shown in SEQ ID No.61 and SEQ ID The primers used to detect CM001598.2:45312379 are the two single-stranded DNAs shown in SEQ ID No.64 and SEQ ID No.65; the primers used to detect CM001598.2:69009397 are the two single-stranded DNAs shown in SEQ ID No.67 and SEQ ID No.68; the primers used to detect CM001600.2:45516740 are the two single-stranded DNAs shown in SEQ ID No.70 and SEQ ID No.71; the primers used to detect CM001601.2:47737780 are the two single-stranded DNAs shown in SEQ ID No.73 and SEQ ID No.74; the primers used to detect CM001602.2:49927473 are the two single-stranded DNAs shown in SEQ ID No.76 and SEQ ID The two single-stranded DNAs are shown in SEQ ID No.77; the primers used to detect the CM001603.2:38707088 are the two single-stranded DNAs shown in SEQ ID No.79 and SEQ ID No.80; the primers used to detect the CM001603.2:46619514 are the two single-stranded DNAs shown in SEQ ID No.82 and SEQ ID No.83; the primers used to detect the CM001606.2:34435705 are the two single-stranded DNAs shown in SEQ ID No.85 and SEQ ID No.86.

Furthermore, the primers for detecting the CM001582.2:86006851 also include the single-stranded DNA shown in SEQ ID No.3; the primers for detecting the CM001583.2:42344562 also include the single-stranded DNA shown in SEQ ID No.6; the primers for detecting the CM001584.2:11650210 also include the single-stranded DNA shown in SEQ ID No.9; the primers for detecting the CM001584.2:12200323 also include the single-stranded DNA shown in SEQ ID No.12; the primers for detecting the CM001584.2:79082958 also include the single-stranded DNA shown in SEQ ID No.15; the primers for detecting the CM001584.2:157466635 also include the single-stranded DNA shown in SEQ ID No.18; the primers for detecting the CM001585.2:5250467 also include the single-stranded DNA shown in SEQ ID The primers for detecting the CM001585.2:30868868 also include the single-stranded DNA shown in SEQ ID No.24; the primers for detecting the CM001588.2:9423474 also include the single-stranded DNA shown in SEQ ID No.27; the primers for detecting the CM001588.2:84941340 also include the single-stranded DNA shown in SEQ ID No.30; the primers for detecting the CM001589.2:75561002 also include the single-stranded DNA shown in SEQ ID No.33; the primers for detecting the CM001590.2:56924859 also include the single-stranded DNA shown in SEQ ID No.36; the primers for detecting the CM001592.2:34421625 also include the single-stranded DNA shown in SEQ ID The primers for detecting CM001592.2:53664048 also include the single-stranded DNA shown in SEQ ID No.42; the primers for detecting CM001592.2:57058734 also include the single-stranded DNA shown in SEQ ID No.45; the primers for detecting CM001593.2:1397868 also include the single-stranded DNA shown in SEQ ID No.48; the primers for detecting CM001594.2:45477903 also include the single-stranded DNA shown in SEQ ID No.51; the primers for detecting CM001594.2:80961940 also include the single-stranded DNA shown in SEQ ID No.54; the primers for detecting CM001595.2:14087769 also include the single-stranded DNA shown in SEQ ID The primers for detecting the CM001595.2:43390933 also include the single-stranded DNA shown in SEQ ID No.60; the primers for detecting the CM001595.2:48968164 also include the single-stranded DNA shown in SEQ ID No.63; the primers for detecting the CM001598.2:45312379 also include the single-stranded DNA shown in SEQ ID No.66; the primers for detecting the CM001598.2:69009397 also include the single-stranded DNA shown in SEQ ID No.69; the primers for detecting the CM001600.2:45516740 also include the single-stranded DNA shown in SEQ ID No.72; the primers for detecting the CM001601.2:47737780 also include the single-stranded DNA shown in SEQ ID The primers for detecting CM001602.2:49927473 also include the single-stranded DNA shown in SEQ ID No.78; the primers for detecting CM001603.2:38707088 also include the single-stranded DNA shown in SEQ ID No.81; the primers for detecting CM001603.2:46619514 also include the single-stranded DNA shown in SEQ ID No.84; the primers for detecting CM001606.2:34435705 also include the single-stranded DNA shown in SEQ ID No.87.

The third is the application of the nucleotide polymorphism at each SNP site in the 11 groups of SNP site combinations mentioned above in the sheep genome (or the nucleotide polymorphism at the 29 SNP sites mentioned above in the sheep genome) in identifying Tan sheep and/or non-Tan sheep.

Fourthly, the application of the nucleotide polymorphism at each SNP site of any one of the 11 groups of SNP site combinations mentioned above in the sheep genome to identify Tan sheep and/or non-Tan sheep.

In a fourth aspect, the present invention claims a set of primers as shown in (B1) or (B2):

(B1) a set of primers for detecting the nucleotides at each SNP site in the 11 groups of SNP site combinations described above in the sheep genome;

(B2) A set of primers for detecting nucleotides at each SNP site in any one of the 11 groups of SNP site combinations described above in the sheep genome.

Among them, the primers used to detect each SNP site are as described above (see Table 2 for details).

The present invention performs whole genome resequencing on 50 Tan sheep and 50 non-Tan sheep to obtain 29 sites, and then analyzes them through R programming to obtain 5-point typing result genotype (identification rate is more than 85%). Then, 1000 Tan sheep and 1000 non-Tan sheep are used for sequenom verification, and the results show that they are consistent with the judgment results. The present invention successfully obtains a combination of SNP sites for identifying Tan sheep and non-Tan sheep, which provides support for the development of Tan sheep variety identification chips, and further provides support for the identification, seed conservation, and genetic breeding of Tan sheep in the future.

DETAILED DESCRIPTION

Unless otherwise specified, the experimental methods used in the following examples are conventional methods.

Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

Example 1: Whole genome resequencing to obtain 5-point genotype results for identifying Tan sheep and non-Tan sheep

Test samples: 50 Tan sheep (from Yanchi County, Ningxia) and 50 non-Tan sheep (from Ningxia, Inner Mongolia and other places).

The blood or ear tissue of the tested sheep was used as a sample to extract genomic DNA, and then the whole genome resequencing method was used to obtain 29 SNP sites, which were then analyzed by R programming to obtain a 5-point typing result genotype (Table 1).

The SNP site selection method is as follows: first mark all sites with P values <-8, and then manually observe whether there are continuous chromosome regions (linkage disequilibrium regions). A continuous region is considered to be a region when there are at least 4 continuous sites. The site selection principle is: 1) First select the most significant site among the sites with P values <-8 in the continuous region. 2) Secondly select the most significant site among the sites with P values <-8 but not in the continuous region; 3) Select the most significant site among the larger continuous regions but without P values <-8.

The whole genome resequencing method is as follows: extract genomic DNA, randomly shear it using Covaris, recover DNA fragments of the required length (0.2-5Kb) by electrophoresis, add adapters, prepare clusters (Illumina HiseqX10), and finally resequence the inserted fragments using the Paired-End (Illumina Hiseq X10) method.

The physical positions of the 29 SNP sites were determined based on the comparison of the standard sequence of the whole genome of Tan sheep, the version number of which is Ovis.Aries V4.0; specifically, as follows:

CM001582.2:86006851 is located at position 86006851 on chromosome 1, and its deoxynucleotide is G or T;

CM001583.2:42344562 is located at position 42344562 on chromosome 2, and its deoxynucleotide is T or C;

CM001584.2:11650210 is located at position 11650210 on chromosome 3, and its deoxynucleotide is T or C;

CM001584.2:12200323 is located at position 12200323 on chromosome 3, and its deoxynucleotide is C or G;

CM001584.2:79082958 is located at position 79082958 on chromosome 3, and its deoxynucleotide is A or G;

CM001584.2:157466635 is located at position 157466635 on chromosome 3, and its deoxynucleotide is C or T;

CM001585.2:5250467 is located at position 5250467 on chromosome 4, and its deoxynucleotide is C or G;

CM001585.2:30868868 is located at position 30868868 on chromosome 4, and its deoxynucleotide is T or C; CM001588.2:9423474 is located at position 9423474 on chromosome 7, and its deoxynucleotide is A or C;

CM001588.2:84941340 is located at position 84941340 on chromosome 7, and its deoxynucleotide is A or C; CM001589.2:75561002 is located at position 75561002 on chromosome 8, and its deoxynucleotide is C or G; CM001590.2:56924859 is located at position 56924859 on chromosome 9, and its deoxynucleotide is A or G; CM001592.2:34421625 is located at position 34421625 on chromosome 11, and its deoxynucleotide is T or C; CM001592.2:53664048 is located at position 53664048 on chromosome 11, and its deoxynucleotide is A or G; C M001592.2:57058734 is located at position 57058734 on chromosome 11, and its deoxynucleotide is C or G; CM001593.2:1397868 is located at position 1397868 on chromosome 12, and its deoxynucleotide is A or G; CM001594.2:45477903 is located at position 45477903 on chromosome 13, and its deoxynucleotide is A or G; CM001594.2:80961940 is located at position 80961940 on chromosome 13, and its deoxynucleotide is A or C; CM001595.2:14087769 is located at position 14087769 on chromosome 14, and its deoxynucleotide is A or T; CM 001595.2:43390933 is located at position 43390933 on chromosome 14, and its deoxynucleotide is A or G; CM001595.2:48968164 is located at position 48968164 on chromosome 14, and its deoxynucleotide is T or G; CM001598.2:45312379 is located at position 45312379 on chromosome 17, and its deoxynucleotide is T or C; CM001598.2:69009397 is located at position 69009397 on chromosome 17, and its deoxynucleotide is A or C; CM001600.2:45516740 is located at position 45516740 on chromosome 19, and its deoxynucleotide is A or C; C M001601.2:47737780 is located at position 47737780 on chromosome 20, and its deoxynucleotide is A or G; CM001602.2:49927473 is located at position 49927473 on chromosome 21, and its deoxynucleotide is T or C; CM001603.2:38707088 is located at position 38707088 on chromosome 22, and its deoxynucleotide is T or C; CM001603.2:46619514 is located at position 46619514 on chromosome 22, and its deoxynucleotide is A or G; CM001606.2:34435705 is located at position 34435705 on chromosome 25, and its deoxynucleotide is C or G;

Table 1 Genotypes of the five typing loci for distinguishing Tan sheep from non-Tan sheep

Note _: The _genotypes of _the five _SNP sites in each group are expressed as _{X1X2_X3X4_X5X6_X7X8_X9X10} , where _{X1 to X10} all represent A or _C or _T or _{G , X1X2} represents the nucleotide at the first SNP site in the group (e.g. AA represents the homozygous nucleotide at the SNP site; AT represents the _heterozygous nucleotide at the SNP site), _X3X4 represents the nucleotide at the second _SNP site in the group, ... _{, X9X10} represents the nucleotide at the fifth SNP site in the group.

Example 2: Verification of the 5-point genotype results of Tan sheep and non-Tan sheep

Verification samples: 1,000 samples of Tan sheep and 1,000 samples of non-Tan sheep.

Genomic DNA was extracted from the blood or ear tissue of the sheep used for verification, and then Sequenom verification was performed.

Among them, the specific steps of Sequenom verification are as follows:

1. Primer Design

Sequenom's Genotyping Tools and MassARRAY Assay Design software were used to design PCR amplification primers and single-base extension primers for the SNP sites to be tested, and then the primers were synthesized by a biological company.

2. DNA quality inspection

The samples to be tested were quantified by spectrophotometer and quality checked by agarose gel electrophoresis. The electrophoresis band of genomic DNA was usually no less than 20 kb. The concentration of DNA that passed the quality inspection was adjusted to 50 ng/μl, transferred to a 96-well plate, and stored at -20°C for future use.

3. Primer dilution

(1) For each SNP, there are three primers (Table 2). The numbers of these three primers are marked on the cap of the primer synthesis tube.

Table 2 Primer information for each SNP

(2) Forward PCR primers and Reverse PCR primers were centrifuged and then water was added. The amount of water added was related to the OD value (both the primer tube and the primer design table indicated). 36 μl of water was added for every 1 OD. After adding water, the mixture was placed at room temperature for 30 min and then vortexed for mixing.

(3) Calculation method for mixing forward PCR primers and reverse PCR primers of multiple SNPs in each well: Take 2.5 μl of each forward PCR primer and reverse PCR primer of each SNP. Add 2.5 μl of water = 500-multiplicity*2.5*2 (multiplicity: number of SNPs in the well)

4. PCR Amplification

PCR amplification was performed using multiplex PCR technology in a 384-well plate, with a total volume of 5 μl for each reaction system.

(1) Prepare PCR master mix solution in a new 2.0 ml EP tube as shown in Table 3.

Table 3 PCR master mix solution

(2) After the prepared PCR master mix solution is shaken and mixed, it is divided into 8 rows of PCR tubes for later use. Using an 8-channel pipette, adjust the sample volume to 4 μL and add the PCR master mix solution to each well of a 384-well plate. The 384-well plate is the PCR reaction plate.

(3) Take out the prepared 96-well plate of DNA samples, use an 8-channel pipette, adjust the sample volume to 1 μL, add it to the corresponding 384 PCR reaction plate, stick a sealing film on it, shake and mix well, and then spin it in the air.

(4) Set the PCR reaction conditions on a 384-well plate compatible PCR instrument as shown in Table 4.

Table 4 PCR reaction conditions

5. Treatment of PCR products with alkaline phosphatase

(1) After the PCR reaction is completed, the 384 reaction plate is removed and spun down.

(2) Prepare alkaline phosphatase treatment reaction solution, SAP Mix, as shown in Table 5.

Table 5 Alkaline phosphatase treatment reaction solution SAP Mix

SAP Mix For each reaction (μL) water 1.53 SAP Buffer (10x) 0.17 SAP enzyme (1.7U/ul) 0.3 Total volume 2

(3) After the prepared SAP mix solution is shaken and mixed, it is divided into 8 rows of PCR tubes for later use. Using an 8-channel pipette, adjust the sample volume to 2 μL and add the SAP Mix to a 384-well PCR reaction plate. Apply a sealing film, shake and mix, and then spin in the air. The total volume of the reaction system is 7 μL (including 5 μL of PCR product and 2 μL of SAP mixture).

(4) Place the 384-well plate on a PCR instrument compatible with 384-well plates and set the PCR reaction conditions as shown in Table 6.

Table 6 PCR reaction conditions

Temperature(℃) Time (minutes) cycle 37 40 1 85 5 1 4 ∞ 1

Start the PCR instrument to carry out alkaline phosphatase treatment reaction.

6. Single base extension

(1) After the alkaline phosphatase treatment, the 384 reaction plate was taken out and spun down, and then a single base extension reaction was performed. The total volume of the reaction system was 9 μl.

(2) Prepare the single base extension reaction solution, EXTEND Mix, as shown in Table 7. (Note: The Extend primer Mix and the Well number must correspond correctly)

Table 7 Single base extension reaction solution EXTEND Mix

EXTEND Mix For each reaction (μL) 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) After the prepared EXTEND mix solution is shaken and mixed, it is divided into 8 rows of PCR tubes for use. Using an 8-channel pipette, adjust the sample volume to 2 μL and add the EXTEND Mix to the 384-well reaction plate. For each reaction well, the single-base extension reaction system contains 7 μL of SAP-treated PCR product and 2 μL of EXTEND Mix solution, with a total system of 9 ul.

(4) Place the 384-well plate on a PCR instrument compatible with 384-well plates and set the PCR reaction conditions as shown in Table 8.

Table 8 PCR reaction conditions

Start the PCR instrument to perform single base extension reaction.

7. Resin Purification

The reaction product was diluted with 16 μl of water and then desalted using a resin.

8. Chip Spotting

The desalted sample is placed on the sample target and allowed to crystallize naturally.

9. Mass spectrometry detection

Perform mass spectrometry detection and collect data.

The results showed that the identification rate of this method was above 85% (meaning: out of 100 sheep to be tested, more than 85 were able to be identified according to this method), as shown in Table 9. The SNP loci combination genotypes of Tan sheep and non-Tan sheep shown in Table 1 were all within the corresponding genotype range shown in Table 1, and their proportions are shown in Table 10.

Table 9 Identification rate of the method of the present invention

Table 10 Sequenom SNP test results of Tan sheep and non-Tan sheep (proportion of genotypes at 5-point typing sites)

Note: The genotype representation method in the table is the same as Table 1.

The results shown in Table 9 indicate that the 5-point genotype results of verifying Tan sheep and non-Tan sheep shown in Table 1 obtained in Example 1 are accurate and reliable. The present invention successfully obtains a combination of SNP sites for identifying Tan sheep and non-Tan sheep, providing support for the identification, seed conservation, and genetic breeding of Tan sheep in the future.

<110> Institute of Animal Science, Ningxia Academy of Agricultural and Forestry Sciences; Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences; Beijing Compson Biotechnology Co., Ltd.

<120> A method for identifying Tan sheep and non-Tan sheep based on genotyping

<130> GNCLN180740

<160> 87

<170> PatentIn version 3.5

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<400> 7

acgttggatg gggtaagtgt gaactccaag 30

<210> 8

<211> 30

<212> DNA

<213> Artificial sequence

<400> 8

acgttggatg tctctcggtg atgtggttgg 30

<210> 9

<211> 15

<212> DNA

<213> Artificial sequence

<400> 9

actccaagag ggaca 15

<210> 10

<211> 30

<212> DNA

<213> Artificial sequence

<400> 10

acgttggatg aaaggcctca gggattgatg 30

<210> 11

<211> 30

<212> DNA

<213> Artificial sequence

<400> 11

acgttggatg aatcagctca gtgtcgctgg 30

<210> 12

<211> 20

<212> DNA

<213> Artificial sequence

<400> 12

ggaatgcgac aggcagcttc 20

<210> 13

<211> 30

<212> DNA

<213> Artificial sequence

<400> 13

acgttggatg tgattcccac tccaatcagg 30

<210> 14

<211> 30

<212> DNA

<213> Artificial sequence

<400> 14

acgttggatg tcatggcatg gttcagagtc 30

<210> 15

<211> 19

<212> DNA

<213> Artificial sequence

<400> 15

gacgcactat acggcacca 19

<210> 16

<211> 30

<212> DNA

<213> Artificial sequence

<400> 16

acgttggatg tgacaaggac taagcaaagc 30

<210> 17

<211> 30

<212> DNA

<213> Artificial sequence

<400> 17

acgttggatg gatggcagcc ctagatcttc 30

<210> 18

<211> 18

<212> DNA

<213> Artificial sequence

<400> 18

cgggtaaaga ttgcaggt 18

<210> 19

<211> 29

<212> DNA

<213> Artificial sequence

<400> 19

acgttggatg tgcaggcgag aactcacag 29

<210> 20

<211> 30

<212> DNA

<213> Artificial sequence

<400> 20

acgttggatg tggtctgccc cagagaaatg 30

<210> 21

<211> 20

<212> DNA

<213> Artificial sequence

<400> 21

cccccatggc cctgtgctgg 20

<210> 22

<211> 30

<212> DNA

<213> Artificial sequence

<400> 22

acgttggatg ttctggttcc aactggtctg 30

<210> 23

<211> 30

<212> DNA

<213> Artificial sequence

<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

<213> Artificial sequence

<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

<212> DNA

<213> Artificial sequence

<400> 28

acgttggatg tgggtcctgg ggagcagcaa 30

<210> 29

<211> 29

<212> DNA

<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

tagatcttcatattttaggttcctt 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

acgttggatgccacagagaaggcaggaaaa 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 taacattggtttccgccagc 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

<212> DNA

<213> Artificial sequence

<400> 83

acgttggatg ttccctgccc ttgatttaag 30

<210> 84

<211> 21

<212> DNA

<213> Artificial sequence

<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

<400> 87

tcagtcctta aacaggaatg ta 22

Claims (11)

1. A method for identifying Tan sheep and non-Tan sheep based on genotyping, comprising the steps of: detecting the nucleotides at each SNP site in 11 groups of SNP site combinations in the genome of the sheep to be tested, determining the The sheep to be tested correspond to the 11 genotypes of the 11 groups of SNP locus combinations; then determine whether the sheep to be tested are Tan sheep as follows: if the sheep to be tested correspond to 11 of the 11 groups of SNP locus combinations Each genotype meets the Tan sheep standard genotype of the corresponding group of SNP locus combinations, then the sheep to be tested is or a candidate for Tan sheep; if the sheep to be tested corresponds to 11 of the 11 groups of SNP locus combinations If the genotypes all conform to the standard genotypes of the non-Tan sheep of the corresponding group of SNP site combinations, then the sheep to be tested is or is a candidate for the non-Tan sheep; The 11 groups of SNP site combinations are as follows (a1)-(a11); the non-Tan sheep standard genotypes and Tan sheep standard genotypes corresponding to the 11 groups of SNP site combinations shown in (a1)-(a11) are sequentially Respectively (b1)-(b11); (a1) Contains 5 SNP sites, namely CM001598.2:69009397, CM001602.2:49927473, CM001603.2:38707088, CM001603.2:46619514 and CM001606.2:34435705; (a2) Contains 5 SNP sites, namely CM001582.2:86006851, CM001598.2:69009397, CM001600.2:45516740, CM001601.2:47737780 and CM001603.2:46619514; (a3) Contains 5 SNP sites, namely CM001582.2:86006851, CM001583.2:42344562, CM001584.2:11650210, CM001584.2:12200323 and CM001588.2:9423474; (a4) Contains 5 SNP sites, namely CM001592.2:57058734, CM001595.2:48968164, CM001598.2:69009397, CM001601.2:47737780 and CM001603.2:38707088; (a5) contains 5 SNP sites, namely CM001585.2:30868868, CM001589.2:75561002, CM001590.2:56924859, CM001598.2:69009397 and CM001606.2:34435705; (a6) contains 5 SNP sites, namely CM001585.2:30868868, CM001588.2:84941340, CM001598.2:45312379, CM001598.2:69009397 and CM001603.2:46619514; (a7) Contains 5 SNP sites, namely CM001585.2:30868868, CM001588.2:84941340, CM001594.2:45477903, CM001594.2:80961940 and CM001603.2:38707088; (a8) Contains 5 SNP sites, namely CM001585.2:30868868, CM001588.2:84941340, CM001593.2:1397868, CM001595.2:14087769 and CM001602.2:49927473; (a9) contains 5 SNP sites, respectively CM001585.2:30868868, CM001588.2:84941340, CM001592.2:34421625, CM001592.2:53664048 and CM001595.2:48968164; (a10) contains 5 SNP sites, namely CM001584.2:79082958, CM001584.2:157466635, CM001601.2:47737780, CM001602.2:49927473 and CM001603.2:38707088; (a11) contains 5 SNP sites, namely CM001584.2:79082958, CM001585.2:5250467, CM001588.2:84941340, CM001595.2:43390933 and CM001595.2:48968164; The physical position of each SNP site in the 11 sets of SNP site combinations is determined based on the comparison of the standard sequence of the whole genome of Tan sheep, and the version number of the standard sequence of the whole genome of Tan sheep is Ovis.Aries V4.0; CM001582.2:86006851 is located at position 86006851 on chromosome 1, and its deoxynucleotide is G or T; CM001583.2:42344562 is located at position 42344562 on chromosome 2, and its deoxynucleotide is T or C; CM001584.2:11650210 is located at position 11650210 on chromosome 3, and its deoxynucleotide is T or C; CM001584.2:12200323 is located at position 12200323 on chromosome 3, and its deoxynucleotide is C or G; CM001584.2:79082958 is located at position 79082958 on chromosome 3, and its deoxynucleotide is A or G; CM001584.2:157466635 is located at position 157466635 on chromosome 3, and its deoxynucleotide is C or T; CM001585.2:5250467 is located at position 5250467 on chromosome 4, and its deoxynucleotide is C or G; CM001585.2:30868868 is located at position 30868868 on chromosome 4, and its deoxynucleotide is T or C; CM001588.2:9423474 is located at position 9423474 on chromosome 7, and its deoxynucleotide is A or C; CM001588.2:84941340 is located at position 84941340 on chromosome 7, and its deoxynucleotide is A or C; CM001589.2:75561002 is located at position 75561002 on chromosome 8, and its deoxynucleotide is C or G; CM001590.2:56924859 is located at position 56924859 on chromosome 9, and its deoxynucleotide is A or G; CM001592.2:34421625 is located at position 34421625 on chromosome 11, and its deoxynucleotide is T or C; CM001592.2:53664048 is located at position 53664048 on chromosome 11, and its deoxynucleotide is A or G; CM001592.2:57058734 is located at position 57058734 on chromosome 11, and its deoxynucleotide is C or G; CM001593.2:1397868 is located at position 1397868 on chromosome 12, and its deoxynucleotide is A or G; CM001594.2:45477903 is located at position 45477903 on chromosome 13, and its deoxynucleotide is A or G; CM001594.2:80961940 is located at position 80961940 on chromosome 13, and its deoxynucleotide is A or C; CM001595.2:14087769 is located at position 14087769 on chromosome 14, and its deoxynucleotide is A or T; CM001595.2:43390933 is located at position 43390933 on chromosome 14, and its deoxynucleotide is A or G; CM001595.2:48968164 is located at position 48968164 on chromosome 14, and its deoxynucleotide is T or G; CM001598.2:45312379 is located at position 45312379 on chromosome 17, and its deoxynucleotide is T or C; CM001598.2:69009397 is located at position 69009397 on chromosome 17, and its deoxynucleotide is A or C; CM001600.2:45516740 is located at position 45516740 on chromosome 19, and its deoxynucleotide is A or C; CM001601.2:47737780 is located at position 47737780 on chromosome 20, and its deoxynucleotide is A or G; CM001602.2:49927473 is located at position 49927473 on chromosome 21, and its deoxynucleotide is T or C; CM001603.2:38707088 is located at position 38707088 on chromosome 22, and its deoxynucleotide is T or C; CM001603.2:46619514 is located at position 46619514 on chromosome 22, and its deoxynucleotide is A or G; CM001606.2:34435705 is located at position 34435705 on chromosome 25, and its deoxynucleotide is C or G; (b1) The standard genotype of non-Tan sheep is any of the following: 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; G; CC_TC_TC_AA_CG; CC_TT_TC_AA_GG; CC_TC_CC_AA_CG; CC_TT_TC_AA_CG; The standard genotype of Tan sheep is any of the following One: 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_GA_GG;AA_CC_TC_AA_CC;CC_CC_TT_GA_GG;AC_CC_CC_AA_CC;AC_CC_TC_GA_CC; (b2) The standard genotype of non-Tan sheep is any of the following: TT_CC_CC_GG_AA; TT_AC_CC_GG_AA; TT_CC_AG_AA; TT_AC_AC_AG_AA; 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; (b3) The standard genotype of non-Tan sheep is any of the following: 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; CC; TT_TC_TT_GG_AC; GT_TT_TT_GG_CC; Tan sheep standard genotype is any of the following: 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; (b4) The standard genotype of non-Tan sheep is any of the following: 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; _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_GG_TC; the standard genotype of Tan sheep is any of the following: CC_GT_AA_AG_TC; GC_GT_AC_GG_CC; GC_GT_AC_AG_TC; CC_GT_AA_AA_CC; _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_GT_AC_AA_CC; CC_GT_AC_AA_TC; CC_TT_AC_AG_CC; (b5) The standard genotype of non-Tan sheep is any of the following: 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; _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; 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; TT_CC_GA_AC_CG; CC_CC_GA_AC_CC; (b6) The standard genotype of non-Tan sheep is any of the following: 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; CC_AA; TC_CC_TC_CC_AA; CC_AA_TT_CC_AA; CC_CC_TC_CC_AA; TC_CA_CC_CC_AA; Tan sheep standard genotype is Any of the following: 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; GA;TT_CC_TT_AA_AA;TC_CC_TC_CC_GA;TT_CC_TC_AC_GA;CC_CA_TT_AC_AA; (b7) The standard genotype of non-Tan sheep is any of the following: 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; 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;Tan sheep standard gene 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; CC_AG_AA_TC; TC_CC_AG_AA_CC; CC_CA_AG_AA_CC; (b8) The standard genotype of non-Tan sheep is any of the following: 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; G_TT_TC; TC_AA_GG_AA_TT; CC_AA_GG_TT_CC; CC_AA_AG_TA_CC; TC_AA_AG_TT_TC; 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; _TT_TC; CC_AA_GG_TA_TC; TC_AA_AG_AA_TT; Tan sheep standard genotype is Any of the following: 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; AA_CC; CC_CA_AA_AA_TC; TT_CC_AA_AA_CC; TT_AA_GG_AA_CC; CC_CA_GG_AA_CC; (b9) The standard genotype of non-Tan sheep is any of the following: 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_CA_TT_AG_TT; TC_CC_TT_GG_TT; CC_CC_TT_AG_GT; TC_CA_TT_AG_TT; The standard genotype of Tan sheep is any of the following One: 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_CC_CT_AG_GT; TT_AA_TT_AA_TT; TT_CC_TT_AA_GT; CC_CA_CT_AG_TT; (b10) Standard genotype of non-Tan sheep is any of the following: 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; _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; 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; 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; (b11) The standard genotype of non-Tan sheep is any of the following: 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; CC AA GG TT; AA GG AA GG TT; AA CC AA GGTT ;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; AG_GT; GA_CC_AA_AA_TT; AA_CC_AA_GG_GT; the standard genotype of Tan sheep is any of the following: AA_GC_AA_GG_GT; AA_GC_CC_AG_GT; GA_GC_CA_AA_GT; GA_GG_CA_AG_TT GG_GC_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; _AG_GT; GA_GC_CC_AG_GT; AA_GG_CC_GG_GT. 2. A method for identifying Tan sheep and non-Tan sheep based on genotyping, comprising the steps of: detecting each SNP site in the SNP site combination shown in claim 1 (a7) in the genome of the sheep to be tested Determine the genotype of the sheep to be tested; then determine whether the sheep to be tested is a Tan sheep as follows: if the genotype of the sheep to be tested meets the (b7) described in claim 1 Tan sheep standard genotype, then the sheep to be tested is or a candidate for Tan sheep; if the genotype of the sheep to be tested meets the non-Tan sheep standard genotype in (b7) described in claim 1, then the The sheep to be tested is or is a candidate for non-Tan sheep. 3. A method for identifying Tan sheep based on genotyping, comprising the steps of: detecting the nucleotides at each SNP site in 11 groups of SNP site combinations in the genome of the sheep to be tested, and determining that the sheep to be tested corresponds to The 11 genotypes of the 11 groups of SNP locus combinations; if the 11 genotypes of the sheep to be tested corresponding to the 11 groups of SNP locus combinations all meet the Tan sheep standard genotypes of the corresponding group of SNP locus combinations, Then the sheep to be tested is or is a candidate for Tan sheep; The 11 groups of SNP locus combinations are (a1)-(a11) described in claim 1; the Tan sheep standard genotypes corresponding to the 11 groups of SNP locus combinations shown in (a1)-(a11) are successively respectively It is the standard genotype of Tan sheep in (b1)-(b11) described in claim 1. 4. A method for identifying Tan sheep based on genotyping, comprising the steps of: detecting the nucleosides at each SNP site in the SNP site combination shown in claim 3 (a7) in the genome of the sheep to be tested acid, determine the genotype of the sheep to be tested; if the genotype of the sheep to be tested meets the standard genotype of Tan sheep in (b7) described in claim 3, then the sheep to be tested is or candidate for Tan sheep sheep. 5. Apply (A1) or (A2) as follows: (A1) The application of the substance used to detect the nucleotides at each SNP site in the 11 groups of SNP site combinations described in claim 1 in the sheep genome in the identification of Tan sheep and/or non-Tan sheep, or in Application in the preparation of a kit for identifying Tan sheep and/or non-Tan sheep; (A2) Application of the substance used to detect the nucleotides at each SNP site in the SNP site combination shown in claim 1 (a7) in the sheep genome to identify Tan sheep and/or non-Tan sheep , or in the preparation of a kit for identifying Tan sheep and/or non-Tan sheep. 6. The application according to claim 5, wherein the substance is a PCR primer and/or a sequencing primer designed for the SNP site to be detected. 7. The application according to claim 6, characterized in that: The primers used to detect the CM001582.2:86006851 are two single-stranded DNAs shown in SEQ ID No.1 and SEQ ID No.2; The primers used to detect the CM001583.2:42344562 are two single-stranded DNAs shown in SEQ ID No.4 and SEQ ID No.5; The primers used to detect the CM001584.2:11650210 are two single-stranded DNAs shown in SEQ ID No.7 and SEQ ID No.8; The primers used to detect the CM001584.2:12200323 are two single-stranded DNAs shown in SEQ ID No.10 and SEQ ID No.11; The primers used to detect the CM001584.2:79082958 are two single-stranded DNAs shown in SEQ ID No.13 and SEQ ID No.14; The primers used to detect the CM001584.2:157466635 are two single-stranded DNAs shown in SEQ ID No.16 and SEQ ID No.17; The primers used to detect the CM001585.2:5250467 are two single-stranded DNAs shown in SEQ ID No.19 and SEQ ID No.20; The primers used to detect the CM001585.2:30868868 are two single-stranded DNAs shown in SEQ ID No.22 and SEQ ID No.23; The primers used to detect the CM001588.2:9423474 are two single-stranded DNAs shown in SEQ ID No.25 and SEQ ID No.26; The primers used to detect the CM001588.2:84941340 are two single-stranded DNAs shown in SEQ ID No.28 and SEQ ID No.29; The primers used to detect the CM001589.2:75561002 are two single-stranded DNAs shown in SEQ ID No.31 and SEQ ID No.32; The primers used to detect the CM001590.2:56924859 are two single-stranded DNAs shown in SEQ ID No.34 and SEQ ID No.35; The primers used to detect the CM001592.2:34421625 are two single-stranded DNAs shown in SEQ ID No.37 and SEQ ID No.38; The primers used to detect the CM001592.2:53664048 are two single-stranded DNAs shown in SEQ ID No.40 and SEQ ID No.41; The primers used to detect the CM001592.2:57058734 are two single-stranded DNAs shown in SEQ ID No.43 and SEQ ID No.44; The primers used to detect the CM001593.2:1397868 are two single-stranded DNAs shown in SEQ ID No.46 and SEQ ID No.47; The primers used to detect the CM001594.2:45477903 are two single-stranded DNAs shown in SEQ ID No.49 and SEQ ID No.50; The primers used to detect the CM001594.2:80961940 are two single-stranded DNAs shown in SEQ ID No.52 and SEQ ID No.53; The primers used to detect the CM001595.2:14087769 are two single-stranded DNAs shown in SEQ ID No.55 and SEQ ID No.56; The primers used to detect the CM001595.2:43390933 are two single-stranded DNAs shown in SEQ ID No.58 and SEQ ID No.59; The primers used to detect the CM001595.2:48968164 are two single-stranded DNAs shown in SEQ ID No.61 and SEQ ID No.62; The primers used to detect the CM001598.2:45312379 are two single-stranded DNAs shown in SEQ ID No.64 and SEQ ID No.65; The primers used to detect the CM001598.2:69009397 are two single-stranded DNAs shown in SEQ ID No.67 and SEQ ID No.68; The primers used to detect the CM001600.2:45516740 are two single-stranded DNAs shown in SEQ ID No.70 and SEQ ID No.71; The primers used to detect the CM001601.2:47737780 are two single-stranded DNAs shown in SEQ ID No.73 and SEQ ID No.74; The primers used to detect the CM001602.2:49927473 are two single-stranded DNAs shown in SEQ ID No.76 and SEQ ID No.77; The primers used to detect the CM001603.2:38707088 are two single-stranded DNAs shown in SEQ ID No.79 and SEQ ID No.80; The primers used to detect the CM001603.2:46619514 are two single-stranded DNAs shown in SEQ ID No.82 and SEQ ID No.83; The primers used to detect the CM001606.2:34435705 are two single-stranded DNAs shown in SEQ ID No.85 and SEQ ID No.86. 8. The application according to claim 7, characterized in that: The primers used to detect the CM001582.2:86006851 also include the single-stranded DNA shown in SEQ ID No.3; The primers used to detect the CM001583.2:42344562 also include the single-stranded DNA shown in SEQ ID No.6; The primers used to detect the CM001584.2:11650210 also include the single-stranded DNA shown in SEQ ID No.9; The primers used to detect the CM001584.2:12200323 also include the single-stranded DNA shown in SEQ ID No.12; The primers used to detect the CM001584.2:79082958 also include the single-stranded DNA shown in SEQ ID No.15; The primers used to detect the CM001584.2:157466635 also include the single-stranded DNA shown in SEQ ID No.18; The primers used to detect the CM001585.2:5250467 also include the single-stranded DNA shown in SEQ ID No.21; The primers used to detect the CM001585.2:30868868 also include the single-stranded DNA shown in SEQ ID No.24; The primers used to detect the CM001588.2:9423474 also include the single-stranded DNA shown in SEQ ID No.27; The primers used to detect the CM001588.2:84941340 also include the single-stranded DNA shown in SEQ ID No.30; The primers used to detect the CM001589.2:75561002 also include the single-stranded DNA shown in SEQ ID No.33; The primers used to detect the CM001590.2:56924859 also include the single-stranded DNA shown in SEQ ID No.36; The primers used to detect the CM001592.2:34421625 also include the single-stranded DNA shown in SEQ ID No.39; The primers used to detect the CM001592.2:53664048 also include the single-stranded DNA shown in SEQ ID No.42; The primers used to detect the CM001592.2:57058734 also include the single-stranded DNA shown in SEQ ID No.45; The primers used to detect the CM001593.2:1397868 also include the single-stranded DNA shown in SEQ ID No.48; The primers used to detect the CM001594.2:45477903 also include the single-stranded DNA shown in SEQ ID No.51; The primers used to detect the CM001594.2:80961940 also include the single-stranded DNA shown in SEQ ID No.54; The primers used to detect the CM001595.2:14087769 also include the single-stranded DNA shown in SEQ ID No.57; The primers used to detect the CM001595.2:43390933 also include the single-stranded DNA shown in SEQ ID No.60; The primers used to detect the CM001595.2:48968164 also include the single-stranded DNA shown in SEQ ID No.63; The primers used to detect the CM001598.2:45312379 also include the single-stranded DNA shown in SEQ ID No.66; The primers used to detect the CM001598.2:69009397 also include the single-stranded DNA shown in SEQ ID No.69; The primers used to detect the CM001600.2:45516740 also include the single-stranded DNA shown in SEQ ID No.72; The primers used to detect the CM001601.2:47737780 also include the single-stranded DNA shown in SEQ ID No.75; The primers used to detect the CM001602.2:49927473 also include the single-stranded DNA shown in SEQ ID No.78; The primers used to detect the CM001603.2:38707088 also include the single-stranded DNA shown in SEQ ID No.81; The primers used to detect the CM001603.2:46619514 also include the single-stranded DNA shown in SEQ ID No.84; The primers used to detect the CM001606.2:34435705 also include the single-stranded DNA shown in SEQ ID No.87. 9. The set of primers is as follows (B1) or (B2): (B1) a set of primers for detecting the nucleotides at each SNP site in the 11 groups of SNP site combinations described in claim 1 in the sheep genome; (B2) a set of primers for detecting the nucleotides at each SNP site in the SNP site combination shown in claim 1 (a7) in the sheep genome; The primers used to detect the CM001582.2:86006851 are two single-stranded DNAs shown in SEQ ID No.1 and SEQ ID No.2; The primers used to detect the CM001583.2:42344562 are two single-stranded DNAs shown in SEQ ID No.4 and SEQ ID No.5; The primers used to detect the CM001584.2:11650210 are two single-stranded DNAs shown in SEQ ID No.7 and SEQ ID No.8; The primers used to detect the CM001584.2:12200323 are two single-stranded DNAs shown in SEQ ID No.10 and SEQ ID No.11; The primers used to detect the CM001584.2:79082958 are two single-stranded DNAs shown in SEQ ID No.13 and SEQ ID No.14; The primers used to detect the CM001584.2:157466635 are two single-stranded DNAs shown in SEQ ID No.16 and SEQ ID No.17; The primers used to detect the CM001585.2:5250467 are two single-stranded DNAs shown in SEQ ID No.19 and SEQ ID No.20; The primers used to detect the CM001585.2:30868868 are two single-stranded DNAs shown in SEQ ID No.22 and SEQ ID No.23; The primers used to detect the CM001588.2:9423474 are two single-stranded DNAs shown in SEQ ID No.25 and SEQ ID No.26; The primers used to detect the CM001588.2:84941340 are two single-stranded DNAs shown in SEQ ID No.28 and SEQ ID No.29; The primers used to detect the CM001589.2:75561002 are two single-stranded DNAs shown in SEQ ID No.31 and SEQ ID No.32; The primers used to detect the CM001590.2:56924859 are two single-stranded DNAs shown in SEQ ID No.34 and SEQ ID No.35; The primers used to detect the CM001592.2:34421625 are two single-stranded DNAs shown in SEQ ID No.37 and SEQ ID No.38; The primers used to detect the CM001592.2:53664048 are two single-stranded DNAs shown in SEQ ID No.40 and SEQ ID No.41; The primers used to detect the CM001592.2:57058734 are two single-stranded DNAs shown in SEQ ID No.43 and SEQ ID No.44; The primers used to detect the CM001593.2:1397868 are two single-stranded DNAs shown in SEQ ID No.46 and SEQ ID No.47; The primers used to detect the CM001594.2:45477903 are two single-stranded DNAs shown in SEQ ID No.49 and SEQ ID No.50; The primers used to detect the CM001594.2:80961940 are two single-stranded DNAs shown in SEQ ID No.52 and SEQ ID No.53; The primers used to detect the CM001595.2:14087769 are two single-stranded DNAs shown in SEQ ID No.55 and SEQ ID No.56; The primers used to detect the CM001595.2:43390933 are two single-stranded DNAs shown in SEQ ID No.58 and SEQ ID No.59; The primers used to detect the CM001595.2:48968164 are two single-stranded DNAs shown in SEQ ID No.61 and SEQ ID No.62; The primers used to detect the CM001598.2:45312379 are two single-stranded DNAs shown in SEQ ID No.64 and SEQ ID No.65; The primers used to detect the CM001598.2:69009397 are two single-stranded DNAs shown in SEQ ID No.67 and SEQ ID No.68; The primers used to detect the CM001600.2:45516740 are two single-stranded DNAs shown in SEQ ID No.70 and SEQ ID No.71; The primers used to detect the CM001601.2:47737780 are two single-stranded DNAs shown in SEQ ID No.73 and SEQ ID No.74; The primers used to detect the CM001602.2:49927473 are two single-stranded DNAs shown in SEQ ID No.76 and SEQ ID No.77; The primers used to detect the CM001603.2:38707088 are two single-stranded DNAs shown in SEQ ID No.79 and SEQ ID No.80; The primers used to detect the CM001603.2:46619514 are two single-stranded DNAs shown in SEQ ID No.82 and SEQ ID No.83; The primers used to detect the CM001606.2:34435705 are two single-stranded DNAs shown in SEQ ID No.85 and SEQ ID No.86. 10. The set of primers according to claim 9, characterized in that: The primers used to detect the CM001582.2:86006851 also include the single-stranded DNA shown in SEQ ID No.3; The primers used to detect the CM001583.2:42344562 also include the single-stranded DNA shown in SEQ ID No.6; The primers used to detect the CM001584.2:11650210 also include the single-stranded DNA shown in SEQ ID No.9; The primers used to detect the CM001584.2:12200323 also include the single-stranded DNA shown in SEQ ID No.12; The primers used to detect the CM001584.2:79082958 also include the single-stranded DNA shown in SEQ ID No.15; The primers used to detect the CM001584.2:157466635 also include the single-stranded DNA shown in SEQ ID No.18; The primers used to detect the CM001585.2:5250467 also include the single-stranded DNA shown in SEQ ID No.21; The primers used to detect the CM001585.2:30868868 also include the single-stranded DNA shown in SEQ ID No.24; The primers used to detect the CM001588.2:9423474 also include the single-stranded DNA shown in SEQ ID No.27; The primers used to detect the CM001588.2:84941340 also include the single-stranded DNA shown in SEQ ID No.30; The primers used to detect the CM001589.2:75561002 also include the single-stranded DNA shown in SEQ ID No.33; The primers used to detect the CM001590.2:56924859 also include the single-stranded DNA shown in SEQ ID No.36; The primers used to detect the CM001592.2:34421625 also include the single-stranded DNA shown in SEQ ID No.39; The primers used to detect the CM001592.2:53664048 also include the single-stranded DNA shown in SEQ ID No.42; The primers used to detect the CM001592.2:57058734 also include the single-stranded DNA shown in SEQ ID No.45; The primers used to detect the CM001593.2:1397868 also include the single-stranded DNA shown in SEQ ID No.48; The primers used to detect the CM001594.2:45477903 also include the single-stranded DNA shown in SEQ ID No.51; The primers used to detect the CM001594.2:80961940 also include the single-stranded DNA shown in SEQ ID No.54; The primers used to detect the CM001595.2:14087769 also include the single-stranded DNA shown in SEQ ID No.57; The primers used to detect the CM001595.2:43390933 also include the single-stranded DNA shown in SEQ ID No.60; The primers used to detect the CM001595.2:48968164 also include the single-stranded DNA shown in SEQ ID No.63; The primers used to detect the CM001598.2:45312379 also include the single-stranded DNA shown in SEQ ID No.66; The primers used to detect the CM001598.2:69009397 also include the single-stranded DNA shown in SEQ ID No.69; The primers used to detect the CM001600.2:45516740 also include the single-stranded DNA shown in SEQ ID No.72; The primers used to detect the CM001601.2:47737780 also include the single-stranded DNA shown in SEQ ID No.75; The primers used to detect the CM001602.2:49927473 also include the single-stranded DNA shown in SEQ ID No.78; The primers used to detect the CM001603.2:38707088 also include the single-stranded DNA shown in SEQ ID No.81; The primers used to detect the CM001603.2:46619514 also include the single-stranded DNA shown in SEQ ID No.84; The primers used to detect the CM001606.2:34435705 also include the single-stranded DNA shown in SEQ ID No.87. 11. Apply (C1) or (C2) as follows: (C1) the application of the nucleotide polymorphism at each SNP site in the 11 groups of SNP site combinations described in claim 1 in the sheep genome to identify Tan sheep and/or non-Tan sheep; (C2) Application of the nucleotide polymorphism at each SNP site in the SNP site combination shown in (a7) in claim 1 in the sheep genome to identify Tan sheep and/or non-Tan sheep .