CN107201409B - SNP marker combination for tracing and identifying beef cattle individual and meat product and application thereof - Google Patents
SNP marker combination for tracing and identifying beef cattle individual and meat product and application thereof Download PDFInfo
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Abstract
The invention discloses an SNP marker combination for tracing and identifying beef cattle individuals and meat products and application thereof. The SNP marker set of the invention comprises 32 highly polymorphic sites distributed in 29 autosomes. The minimum allele frequency of the combined locus is more than 0.35. Experiments prove that: the combination can be successfully applied to individual identification and meat tracing of beef cattle except yak varieties in the market, is favorable for realizing full chain tracing of breeding, slaughtering and selling, and really ensures the safety of food.
Description
Technical Field
The invention belongs to the field of individual identification and traceability detection, and particularly relates to an SNP marker combination for beef cattle individual and meat traceability identification and application thereof.
Background
The food tracing technology is considered as a means for ensuring food safety and protecting the legitimate rights and interests of consumers to be most trustable. At present, the traditional electronic ear tag technology (radio frequency identification system) has the problems of easy loss, easy tampering of traceability information, difficult realization of traceability of meat products to livestock and cattle breeding conditions and the like. However, the gene tracing technology represented by ssr (simple sequence repeat) and snp (single nucleotide polymorphism) has the advantages of uniqueness, irredifiable modification, difficult loss and the like, and is gradually becoming a powerful supplement to the electronic ear tag tracing system. The gene tracing technology is applied to a meat food tracing management system, so that the full-chain tracing of cultivation, slaughtering and sale is realized, and the safety of food is really guaranteed.
SNP is used as a third-generation molecular marker technology, and has the advantages of abundant quantity, stable heredity and low type judgment error rate compared with a microsatellite (SSR). And at present, the method is rich in SNP typing methods, such as a TaqMan probe method, a high resolution melting curve (HRM), second generation sequencing (NGS), mass spectrometry (MALDI TOF), a DNA chip and the like, and the methods are convenient to use, high in automation degree and convenient for SNP tracing application.
Disclosure of Invention
An object of the present invention is to provide a novel use of a substance for detecting X;
the X is at least one of the following SNP sites: rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs 18429002, rs29025314, rs29025593, rs29012633, rs 29019919957, rs29010153, rs 29022322377, rs29010275, rs29001970, rs29009713, rs 29021821886, and rs 29011378.
The present invention provides use of a substance for detecting X in any one of the following 1) to 6):
1) tracing and identifying beef products;
2) preparing a beef product tracing and identifying product;
3) distinguishing yak meat from other kinds of beef;
4) preparing products for distinguishing yak meat and other varieties of beef;
5) identifying a cattle individual;
6) preparing a product for individual identification of cattle;
in the application, the X is the following X1) or X2) or X3):
x1) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, and rs 29014143;
x2) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs17871566, and rs 29014143;
x3) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, rs29012633, rs29019957, rs29010153, rs 29022322377, rs29010275, rs29001970, rs29009713, rs29021886, and rs 29011378.
In the above-mentioned application, the first and second substrates,
the substance for detecting rs29019900 consists of a single-stranded DNA molecule shown in a sequence 1 and a single-stranded DNA molecule shown in a sequence 2;
the substance for detecting rs29010374 consists of a single-stranded DNA molecule shown in sequence 3 and a single-stranded DNA molecule shown in sequence 4;
the substance for detecting rs29016185 consists of a single-stranded DNA molecule shown in a sequence 5 and a single-stranded DNA molecule shown in a sequence 6;
the substance for detecting rs29010006 consists of a single-stranded DNA molecule shown in a sequence 7 and a single-stranded DNA molecule shown in a sequence 8;
the substance for detecting rs29010035 consists of a single-stranded DNA molecule shown in sequence 9 and a single-stranded DNA molecule shown in sequence 10;
the substance for detecting rs29023422 consists of a single-stranded DNA molecule shown in a sequence 11 and a single-stranded DNA molecule shown in a sequence 12;
the substance for detecting rs29014752 consists of a single-stranded DNA molecule shown in sequence 13 and a single-stranded DNA molecule shown in sequence 14;
the substance for detecting rs17871566 consists of a single-stranded DNA molecule shown in sequence 15 and a single-stranded DNA molecule shown in sequence 16;
the substance for detecting rs29014143 consists of a single-stranded DNA molecule shown in a sequence 17 and a single-stranded DNA molecule shown in a sequence 18;
the substance for detecting rs29014953 consists of a single-stranded DNA molecule shown in sequence 19 and a single-stranded DNA molecule shown in sequence 20;
the substance for detecting rs17870274 consists of a single-stranded DNA molecule shown in sequence 21 and a single-stranded DNA molecule shown in sequence 22;
the substance for detecting rs29021631 consists of a single-stranded DNA molecule shown in sequence 23 and a single-stranded DNA molecule shown in sequence 24;
the substance for detecting rs29024289 consists of a single-stranded DNA molecule shown in a sequence 25 and a single-stranded DNA molecule shown in a sequence 26;
the substance for detecting rs29017633 consists of a single-stranded DNA molecule shown in sequence 27 and a single-stranded DNA molecule shown in sequence 28;
the substance for detecting rs41255624 consists of a single-stranded DNA molecule shown in sequence 29 and a single-stranded DNA molecule shown in sequence 30;
the substance for detecting rs29019421 consists of a single-stranded DNA molecule shown as a sequence 31 and a single-stranded DNA molecule shown as a sequence 32;
the substance for detecting rs29023295 consists of a single-stranded DNA molecule shown in a sequence 33 and a single-stranded DNA molecule shown in a sequence 34;
the substance for detecting rs29020547 consists of a single-stranded DNA molecule shown in a sequence 35 and a single-stranded DNA molecule shown in a sequence 36;
the substance for detecting rs29009782 consists of a single-stranded DNA molecule shown as a sequence 37 and a single-stranded DNA molecule shown as a sequence 38;
the substance for detecting rs29020876 consists of a single-stranded DNA molecule shown in sequence 39 and a single-stranded DNA molecule shown in sequence 40;
the substance for detecting rs29018402 consists of a single-stranded DNA molecule shown in a sequence 41 and a single-stranded DNA molecule shown in a sequence 42;
the substance for detecting rs29025314 consists of a single-stranded DNA molecule shown in sequence 43 and a single-stranded DNA molecule shown in sequence 44;
the substance for detecting rs29025593 consists of a single-stranded DNA molecule shown in a sequence 45 and a single-stranded DNA molecule shown in a sequence 46;
the substance for detecting rs29012633 consists of a single-stranded DNA molecule shown in sequence 47 and a single-stranded DNA molecule shown in sequence 48;
the substance for detecting rs29019957 consists of a single-stranded DNA molecule shown in sequence 49 and a single-stranded DNA molecule shown in sequence 50;
the substance for detecting rs29010153 consists of a single-stranded DNA molecule shown in a sequence 51 and a single-stranded DNA molecule shown in a sequence 52;
the substance for detecting rs29022377 consists of a single-stranded DNA molecule shown in sequence 53 and a single-stranded DNA molecule shown in sequence 54;
the substance for detecting rs29010275 consists of a single-stranded DNA molecule shown in sequence 55 and a single-stranded DNA molecule shown in sequence 56;
the substance for detecting rs29001970 consists of a single-stranded DNA molecule shown in a sequence 57 and a single-stranded DNA molecule shown in a sequence 58;
the substance for detecting rs29009713 consists of a single-stranded DNA molecule shown in sequence 59 and a single-stranded DNA molecule shown in sequence 60;
the substance for detecting rs29021886 consists of a single-stranded DNA molecule shown in sequence 61 and a single-stranded DNA molecule shown in sequence 62;
and the substance for detecting rs29011378 consists of a single-stranded DNA molecule shown in sequence 63 and a single-stranded DNA molecule shown in sequence 64.
In practical application, the SNP sites and the number of the SNP sites can be finally determined according to the variety composition of the target population, the total population number, the required accuracy and the acceptable cost. As the number of the selected SNP sites increases, the number of the individuals which can be identified is larger, and when the number of the groups is fixed, the more the number of the SNP sites is used, the more accurate the individual distinguishing and identifying effect is.
Another object of the present invention is to provide a method for tracing and identifying beef products.
The beef product traceability identification method provided by the invention comprises the following steps: carrying out genotype identification on each cattle to be listed in the cattle breeding land by adopting the substances to obtain the genotype of each cattle to be listed; when the cattle are slaughtered and marketed, taking a meat sample of the cattle, and carrying out genotype identification on the meat sample by adopting the substance to obtain the genotype of the meat sample; if the genotype of the meat sample is the same as the genotype of a certain cattle to be marketed, the meat sample is derived from the cattle.
In the method, the substance is a primer set A or a primer set B;
the primer set A consists of a substance for detecting the rs29019900, a substance for detecting the rs29010374, a substance for detecting the rs29016185, a substance for detecting the rs29010006, a substance for detecting the rs29010035, a substance for detecting the rs29023422, a substance for detecting the rs29014752, a substance for detecting the rs17871566 and a substance for detecting the rs29014143 in claim 3;
the set of primers B consists of the substance for detecting rs29019900, the substance for detecting rs29010374, the substance for detecting rs29016185, the substance for detecting rs29010006, the substance for detecting rs29010035, the substance for detecting rs29023422, the substance for detecting rs29014752, the substance for detecting rs17871566, the substance for detecting rs29014143, the substance for detecting rs29014953, the substance for detecting rs17870274, the substance for detecting rs29021631, the substance for detecting rs29024289, the substance for detecting rs29017633, the substance for detecting rs 415624, the substance for detecting rs 29019419421, the substance for detecting rs29023295, the substance for detecting rs29020547, the substance for detecting rs29009782, the substance for detecting rs29020876, the substance for detecting rs 18402, the substance for detecting rs29025314, the substance for detecting rs29025593, the substance for detecting rs 290255957, the substance for detecting rs 29029029029010133, the substance for detecting rs 2905953, the substance for detecting rs 29022322322322322322322322322322377, the substance for detecting rs 2905953, the substance for detecting rs 2902902905953, the substance for detecting rs 2905953, the substance for detecting, A substance to detect the rs29010275, a substance to detect the rs29001970, a substance to detect the rs29009713, a substance to detect the rs29021886, and a substance composition to detect the rs 29011378.
In practical application, the source of the meat sample can be judged by detecting the genotype barcode of each blood sample of the cattle to be marketed in the cattle breeding area and the slaughtered and marketed meat sample. The specific compiling method of the genotype bar code is as follows: the genotype barcode of each sample sequentially consists of a genotype barcode at a site rs29019900, a genotype barcode at an rs29010374, a genotype barcode at an rs29016185, a genotype barcode at an rs29010006, a genotype barcode at an rs29010035, a genotype barcode at an rs29023422, a genotype barcode at an rs29014752, a genotype barcode at an rs17871566 and a genotype barcode at an rs29014143, the genotype barcode at the site rs29019900 is taken as an example to illustrate the compiling method of the genotype barcode at each site, the mutation type at the site rs29019900 is A/G, the genotype of a sample in which the rs29019900 sites of two homologous chromosomes are A is taken as AA, the genotype of a sample in which the rs29019900 sites of two homologous chromosomes are G is taken as GG, and the genotype of a sample in which the rs29019900 sites of two homologous chromosomes are A and G is taken as AG. When the genotype of the sample at the rs29019900 site is AA, the genotype barcode of the sample at the rs29019900 site is marked as '0'; when the genotype of the sample at the rs29019900 site is AG, the genotype barcode of the sample at the rs29019900 site is marked as "1"; when the genotype of the sample at the rs29019900 site is GG, the genotype barcode of the sample at the rs29019900 site is marked as "2". By analogy, the genotype barcode of the sample at other 8 SNP sites is obtained respectively, and the genotype barcodes at each SNP site are arranged in sequence, and finally the genotype barcode of the sample at the 9 SNP sites is obtained. And obtaining respective genotype bar codes after each sample is subjected to typing measurement, and judging the source of the sample by comparing the bar codes. If the genotype bar codes of a certain meat sample and a certain blood sample are completely consistent, the meat sample and the blood sample are from the same individual (except for the individuals of the same ovum twins), all the recorded information of the corresponding cattle individual can be known according to the source of the blood sample, and the purpose of tracing is achieved.
In practical application, any one or two or three or more of the 32 SNP sites in table 3 may be selected as a SNP site combination for beef cattle product traceability identification according to actual needs, or all 32 SNP sites may be selected as a SNP site combination for beef cattle product traceability identification. In the specific embodiment of the invention, 9 SNP loci and all 32 SNP loci are respectively selected for carrying out the source tracing identification of beef cattle products. The identification result shows that: no matter all 32 SNP loci or part of 32 SNP loci are selected for beef cattle product traceability identification, the whole chain traceability information of beef cattle in the market can be established, and meat products pretending to be specific brands can be identified and further used for identifying authenticity of identification.
The invention also aims to provide a method for distinguishing the yak meat from other varieties of beef.
The method for distinguishing the yak meat from other varieties of beef provided by the invention comprises the following steps: carrying out genotype identification on the beef to be detected by adopting the substances to obtain the genotype of the beef to be detected;
if the genotype of the beef to be detected at each selected SNP site is homozygous, the beef to be detected is yak meat or is a candidate;
if the genotype of the beef to be detected at each selected SNP site is not homozygous, the beef to be detected is or is selected as non-yak beef.
In the method, the substance is a primer set C; the primer set C consists of a substance for detecting the rs29019900, a substance for detecting the rs29010374, a substance for detecting the rs29016185, a substance for detecting the rs29010006, a substance for detecting the rs29010035, a substance for detecting the rs29023422, a substance for detecting the rs17871566 and a substance for detecting the rs 29014143.
In the method, if the bases of two homologous chromosomes of the beef to be detected at a certain SNP locus are the same, the genotype of the beef to be detected at the SNP locus is homozygous.
In practical application, the site having no polymorphism in yak population and higher polymorphism in other populations in table 3, i.e. one or two or more of the other 30 SNP sites except the rs29014752 site and the rs29020876 site, can be selected for distinguishing yak meat from other varieties of beef. Whether the genotypes of the beef samples to be detected at the selected SNP sites are homozygous or not can be judged by determining the genotype barcode of the beef samples to be detected at the selected SNP sites according to the method. If the genotype barcode of each SNP locus in the genotype barcode of a certain beef sample to be detected is an even number, the sample to be detected is yak meat or is a candidate; if the genotype bar codes of all SNP sites in the genotype bar code of a certain beef sample to be detected are not even numbers, the beef to be detected is or is selected as non-yak meat.
In the above method, the genotyping is based on a high-throughput Hi-SNP typing platform.
The invention finally aims to provide a product for tracing and identifying beef products or distinguishing yak meat from other varieties of beef or individual identification of the beef.
The product for tracing and identifying the beef product or distinguishing the yak meat from other varieties of beef or individual identification of the beef provided by the invention is any one of the following products (1) to (5):
(1) the primer set A or the primer set B;
(2) the primer set C;
(3) the substance for detecting X described above;
(4) a PCR reagent containing the primer set A or the primer set B or the primer set C or the substance for detecting X;
(5) a kit comprising the primer set A or the primer set B or the primer set C or the substance for detecting X.
In the above application or the above method or the above product, the beef cattle are beef cattle, specifically simmental cattle, russian cattle, Xinjiang black cattle, lizhan cattle, south yang yellow cattle, black and cattle or red cattle in Jiaxian county.
The invention has the following advantages;
(1) the selected polymorphic sites are rich, and the total number is 32;
(2) the selected loci have high polymorphism, and the minimum allele frequency is more than 0.35;
(3) the markers selected in the present invention are not linked: all 32 loci are distributed on 29 autosomes, the minimum distance between loci on the same chromosome is 1.7Mb, the maximum distance between loci on the same chromosome is 6.3Mb, and the possibility of linkage disequilibrium is extremely low;
(4) the marker combination selected by the invention can be used for individual identification and meat source tracing of other groups except yak groups in the market in China.
The invention provides a group of SNP marker combinations for individual identification and meat source tracing of beef cattle in Chinese market. This combination contains 32 highly polymorphic sites distributed over 29 autosomes. The minimum allele frequency of the combined locus is more than 0.35. The combination can be successfully applied to individual identification and meat tracing of beef cattle except yak varieties in the market, is favorable for realizing full chain tracing of breeding, slaughtering and selling, and really ensures the safety of food.
Detailed Description
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
In the quantitative tests in the following examples, three replicates were set up and the results averaged.
Example 1 SNP marker combination for tracing and identifying beef cattle individuals and meat products
Firstly, collecting samples and extracting genome DNA
1. Collection of samples
The samples of the present invention were obtained from 192 individuals of 8 species on the chinese market, simmental cattle (n-43), luxi yellow cattle (n-34), xinjiang black cattle (n-24), lizardn cattle (n-30), yak (n-16), south yang yellow cattle (n-15), black and cattle (n-15), and zhou red cattle (n-15). A large population of 192 individuals of 8 varieties is a mixed population.
2. Extraction of genomic DNA
Blood samples were collected using EDTA anticoagulant blood collection tubes (5mL), and blood DNA was extracted using the GeneJET Genomic DNA purification Kit (Thermo Scientific, # k 0721). The purity and concentration of DNA are determined by using Nanodrop2000c, the integrity of DNA fragments is inspected by using 0.8% agarose gel electrophoresis, and the DNA meeting the requirements is stored at the temperature of-20 ℃ for later use.
Second, screening of SNP site
1. SNP site selection and primer design
Firstly, 60 SNP loci are selected from NCBI dbSNP database (www.ncbi.nlm.nih.gov/projects/SNP /), the loci are distributed on 29 chromosomes of a bovine genome, each chromosome has about 1-3 loci, the distance between adjacent loci on the same chromosome is more than 1Mb, most loci are positioned in gene intron regions, and most loci have reference gene frequency and genotype frequency, so that the linkage correspondence between loci can be avoided as much as possible, and the efficiency of screening the loci can be improved. The information and primers for position 60 are shown in Table 1.
TABLE 1, 60 polymorphic sites and primer information
2. Detection of SNP polymorphism and polymorphism analysis
The high-throughput Hi-SNP typing platform is adopted to detect SNP polymorphism, and three rounds of multiplex PCR are combined with a second-generation sequencing technology to detect the genotypes of 192 individuals at each site. The observed heterozygosity (HObs), expected heterozygosity (Hexp), Polymorphic Information Content (PIC) and Minimum Allele Frequency (MAF) were counted for each site. Sites with MAF <0.35 were truncated to ensure site identification discrimination efficiency. Finally, 32 polymorphic loci are obtained, and the specific information is shown in Table 2.
TABLE 2, 32 polymorphism site polymorphisms
Application of three, 32 SNP loci in bovine individual identification
1. Coupling probability of 32 SNP sites in mixed population and each variety
The coupling probability (MP) refers to the probability that two random individuals have the same genotype. The value reflects the discrimination identification potency of the locus and its combination for the individuals, and the smaller the value, the higher the discrimination potency between the individuals, and conversely, the worse the discrimination effect.
The MP values of the 32 SNP sites in all individuals and different varieties are shown in Table 3. First, it can be seen that the MP values of the SNP sites are slightly different between different populations and varieties, for example, the MP values of the SNP sites in the mixed population are between 0.334 and 0.389, and the average value is 0.359. The values increased slightly between the different breeds, such as between 0.334 and 0.463 with a mean value of 0.387 in the Simmental cattle breed and between 0.337 and 0.531 with a mean value of 0.403 in the Xinjiang black cattle. The MP value of each SNP locus differs in value among different varieties, which is related to the polymorphism condition of the locus in different populations, and the difference of the genetic background of the different populations is also indicated.
In addition, it can be seen from table 3 that: most of the 32 loci do not show polymorphism in the yak population, and only rs29014752 and rs29020876 show certain polymorphism. This shows that the genetic structure of the yak group is different from the genetic structures of other beef cattle groups, so the identification traceability site aiming at the yak group has pertinence.
TABLE 3 coupling probability of 32 SNPs sites in mixed population and different varieties
2. Individual identification effects of different numbers of SNP site combinations in different populations
The coupling probability of the SNP locus combinations with different numbers in the mixed population and each breed (except yak) is respectively calculated, the coupling probability is totally divided into 12 groups, the number of the SNPs in each group is respectively 5, 9, 11, 14, 16, 18, 20, 23, 25, 27, 29 and 32, and each group comprises the locus with the highest polymorphism in each population. Wherein the SNP site combinations of 5 in number are composed of the SNP sites represented by the sequence numbers 1 to 5 in Table 3, the SNP site combinations of 9 in number are composed of the SNP sites represented by the sequence numbers 1 to 9 in Table 3, the SNP site combinations of 11 in number are composed of the SNP sites represented by the sequence numbers 1 to 11 in Table 3, and so on.
The results are shown in Table 4. The first set of 5 SNP site combinations, the discrimination potency in each population ranged from 4.4 thousandths (Luxi cattle) to 5.9 thousandths (Nanyang cattle). That is, when 5 SNP site combinations are used, the number of distinguishable individuals is about 170-. With the continuous increase of the number of SNP loci, the MP value is remarkably reduced, and the probability that two random individuals have the same genotype is gradually reduced. When the number of SNP sites is increased to 9, the other groups except the Nanyang cattle group can reach about 6 parts per million. When the number of SNP sites increased to 14, the MP value decreased to 3.8 parts per million. When the number of SNP sites increases to 32, taking the mixed population as an example, the probability that two random individuals have the same genotype is 5.616E-15, i.e., about 5.6 individuals in a trillion population may have the same genotype. Therefore, with the increase of the number of the SNP sites, the number of the individuals which can be identified is larger, when the population quantity is fixed, the number of the SNP sites which are used is larger, the effect of individual distinguishing and identifying is more accurate, and in practical application, which SNP sites and the number of the SNP sites which are used can be finally determined according to the variety composition of the target population, the total population quantity, the required accuracy and the acceptable cost.
TABLE 4 coupling probability of different SNP site combinations in Mixed populations and varieties
Example 2 application of SNP site combination in beef cattle product traceability identification
1. Sample collection and genomic DNA extraction
In a certain beef cattle breeding base (M), 15 cattle blood samples including Simmental cattle, Luxi yellow cattle and Xinjiang black cattle are randomly collected in a tail vein blood sampling mode, the number of the blood sample is MB1-MB15, and the ear tag number of each cattle is recorded. And taking corresponding meat samples with the meat sample number of MM1-MM15 when the cattle are slaughtered and marketed, and randomly collecting meat samples of other 5 cattle on the market with the meat sample number of NM1-NM 5. DNA extraction and quality control were the same as in example 1.
2. Genotype determination and meat tracing identification of sample
(1)9 SNP site combinations
The following 9 SNP sites with highest individual discrimination efficiency were used: rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566 and rs29014143, all blood and meat samples were typed, the genotype of each sample was counted, and the genotype barcodes were compiled for alignment. The specific compiling method of the genotype bar code comprises the following steps: the genotype barcode of each sample sequentially consists of a genotype barcode at a site rs29019900, a genotype barcode at an rs29010374, a genotype barcode at an rs29016185, a genotype barcode at an rs29010006, a genotype barcode at an rs29010035, a genotype barcode at an rs29023422, a genotype barcode at an rs29014752, a genotype barcode at an rs17871566 and a genotype barcode at an rs29014143, the genotype barcode at the site rs29019900 is taken as an example to illustrate the compiling method of the genotype barcode at each site, the mutation type at the site rs29019900 is A/G, the genotype of a sample in which the rs29019900 sites of two homologous chromosomes are A is taken as AA, the genotype of a sample in which the rs29019900 sites of two homologous chromosomes are G is taken as GG, and the genotype of a sample in which the rs 29000 sites of two homologous chromosomes are A and G is taken as AG. When the genotype of the sample at the rs29019900 site is AA, the genotype barcode of the sample at the rs29019900 site is marked as '0'; when the genotype of the sample at the rs29019900 site is AG, the genotype barcode of the sample at the rs29019900 site is marked as "1"; when the genotype of the sample at the rs29019900 site is GG, the genotype barcode of the sample at the rs29019900 site is marked as "2". By analogy, the genotype barcode of the sample at other 8 SNP sites is obtained respectively, and the genotype barcodes at each SNP site are arranged in sequence, and finally the genotype barcode of the sample at the 9 SNP sites is obtained. And obtaining respective genotype bar codes after each sample is subjected to typing measurement, and judging the source of the sample by comparing the bar codes. If the genotype bar codes of a certain meat sample and a certain blood sample are completely consistent, the meat sample and the blood sample are from the same individual (except for the individuals of the same ovum twins), all the recorded information of the corresponding cattle individual can be known according to the source of the blood sample, and the purpose of tracing is achieved.
The results are shown in Table 5. As can be seen from table 5, the corresponding blood samples of the meat samples from the beef cattle breeding base (M) can be found through the genotype barcode, and the corresponding individual cattle can be found. While the other 5 meat samples were randomly collected on the market without having the same genotype barcode, indicating that they did not come from the beef breeding base (M).
TABLE 5 genotype barcode comparison of traceable samples
| Meat sample | Animal ear tag | Genotype bar code | Results |
| MM1 | 743692 | 020222102 | In agreement with MB1 |
| MM2 | 357900 | 002121010 | In agreement with MB2 |
| MM3 | 357964 | 002121122 | In agreement with MB3 |
| MM4 | 22649 | 211001110 | In agreement with MB4 |
| MM5 | 207487 | 011012221 | In agreement with MB5 |
| MM6 | 210326 | 012012020 | In agreement with MB6 |
| MM7 | KD—BC3 | 020112112 | In agreement with MB7 |
| MM8 | 173900 | 020212120 | In agreement with MB8 |
| MM9 | 173143 | 122222011 | In agreement with MB9 |
| MM10 | 138 | 111211011 | In agreement with MB10 |
| MM11 | Zf--c3 | 121212212 | In agreement with MB11 |
| MM12 | Zf--c6 | 121202222 | In agreement with MB12 |
| MM13 | 743719 | 221000121 | In agreement with MB13 |
| MM14 | 357944 | 122202102 | In agreement with MB14 |
| MM15 | 357951 | 112102102 | In agreement with MB15 |
| NMM1 | No information | 010102202 | Non-uniform sample |
| NMM2 | No information | 021121122 | Non-uniform sample |
| NMM3 | No information | 111212202 | Non-uniform sample |
| NMM4 | No information | 111012002 | Non-uniform sample |
| NMM5 | No information | 020021221 | Non-uniform sample |
(2)32 SNP site combinations
All the 32 SNP sites in the table 3 are adopted to carry out typing determination on all blood and meat samples according to the method in the step (1), the genotype of each sample is counted, and a genotype barcode is coded for comparison. The results show that: the meat samples from the beef cattle breeding base (M) can also respectively find the corresponding blood samples through the genotype bar codes, and then find the corresponding cattle individuals. While the other 5 meat samples were randomly collected on the market without having the same genotype barcode, indicating that they did not come from the beef breeding base (M).
Therefore, no matter all 32 SNP loci are selected for beef product traceability identification or part of 32 SNP loci are selected for beef product traceability identification, the full-chain traceability information of beef cattle in the market can be established, and the beef products pretending to be specific brands can be identified and further used for identifying the authenticity of the identification.
In practical application, meat source tracing identification can be carried out according to the following method, namely how to determine a cattle individual from which a certain beef comes on the market:
selecting at least one of the following SNP sites: rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs 18429002, rs29025314, rs29025593, rs29012633, rs 29019919957, rs29010153, rs 29022322377, rs29010275, rs29001970, rs29009713, rs 29021821886 and rs 11378, and performing genotype identification on each cattle blood sample to be taken in a cattle breeding place according to the selected SNP locus to obtain the genotype of each cattle to be taken in the market; when the cattle in the cattle breeding place are slaughtered and marketed, taking a meat sample, and carrying out genotype identification on the meat sample by adopting the selected SNP locus to obtain the genotype of the meat sample; if the genotype of the meat sample is the same as the genotype of a blood sample of a bovine to be marketed, the meat sample is derived from the bovine.
Example 3 application of SNP locus combination in identification of yak meat and other varieties of beef
1. Sample collection and genomic DNA extraction
The method is characterized in that 36 yak meat samples from Qinghai, Gansu, Tibet, Yunnan and the like are respectively collected, 50 beef samples of other varieties including Simmental cattle, Luxi yellow cattle, Xinjiang black cattle and Limangzan cattle are respectively subjected to DNA extraction and quality control, and the specific operation is the same as that in example 1.
2. Genotype determination of sample and identification of yak meat and other varieties of beef
Selecting sites (other 30 SNP sites except rs29014752 and rs29020876 sites) which have no polymorphism in the yak group and have higher polymorphism in other groups in the table 3, and selecting the following 8 SNP sites for identifying the yak meat and other varieties of beef from the 30 SNP sites: rs2901990, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs17871566 and rs29014143, all beef samples were genotyped using the selected combination of SNP sites, the genotype of each sample was counted and the genotype barcodes were compiled for alignment. The specific construction method of the genotype barcode was the same as that in example 2.
The results of the partial detection are shown in Table 6. As can be seen from the table: the genotype bar codes of all SNP sites in the genotype bar codes of the yak meat samples are even numbers, namely the genotype of the yak meat samples at all the selected SNP sites is homozygous; and the genotype bar codes of all SNP sites in the genotype bar codes of the non-yak meat samples are uneven numbers, namely the genotype of the non-yak meat samples at all the selected SNP sites is uneven and homozygous. Therefore, whether the beef to be detected is yak meat or not can be judged by detecting the genotype of the beef to be detected based on the SNP locus combination.
TABLE 6 genotype bar codes of yak meat and other beef varieties
Therefore, in the actual detection, whether the beef to be detected is yak meat can be detected or detected in an auxiliary mode according to the following method:
selecting at least one of the following SNP sites: rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29018402, rs29025314, rs29025593, rs29012633, rs29019957, rs29010153, rs29022377, rs29010275, rs29001970, rs29009713, rs 29021821821886 and rs29011378, detecting the genotype of the beef to be detected at each SNP site according to the selected SNP site, and judging whether the beef to be detected is yak beef according to the genotype of each SNP site:
if the genotype of the beef to be detected at each selected SNP site is homozygous, the beef to be detected is yak meat or is a candidate;
if the genotype of the beef to be detected at each selected SNP site is not homozygous, the beef to be detected is or is selected as non-yak beef.
Sequence listing
<110> institute of agricultural quality standards and testing technology of Chinese academy of agricultural sciences
<120> SNP marker combination for tracing and identifying beef cattle individual and meat product and application thereof
<160> 64
<210> 1
<211> 18bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 1
acttgctgcc agggaagg 18
<210> 2
<211> 24bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 2
aattcagtgt gatttcactc tccc 24
<210> 3
<211> 30bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 3
ttcacgtata ctttctcaat ttatacaatg 30
<210> 4
<211> 21bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 4
ggaagagcac gtgacatttg c 21
<210> 5
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 5
ctttctcctc tactttcttc ttcatg 26
<210> 6
<211> 25bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 6
ggctacaacc aggatattga agtac 25
<210> 7
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 7
gacattttag aagttagcct tgagtg 26
<210> 8
<211> 23bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 8
caatggatga gctctttaaa agg 23
<210> 9
<211> 28bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 9
caaaggaaat agacttttta aagtcttg 28
<210> 10
<211> 20bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 10
ctgagtctgg cctgggtgta 20
<210> 11
<211> 22bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 11
tgtagaaatc aagtggcctg ga 22
<210> 12
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 12
caggttttct tttaacagaa aaagac 26
<210> 13
<211> 22bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 13
cacctctgaa ggtcaccaat ca 22
<210> 14
<211> 22bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 14
gcactacaaa gaaggatcct gc 22
<210> 15
<211> 21bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 15
agatgatcag gacagccaag g 21
<210> 16
<211> 23bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 16
gtgctcctga atcatcagct att 23
<210> 17
<211> 20bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 17
catgaaagga agcaacccag 20
<210> 18
<211> 20bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 18
cagctgagct tcctgggaag 20
<210> 19
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 19
cactatatta aatctttgct gccaag 26
<210> 20
<211> 21bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 20
agtacagacc agatggccga c 21
<210> 21
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 21
gccttcacta ttcttaagtc cttttg 26
<210> 22
<211> 24bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 22
gatcaaggca gaactacact accc 24
<210> 23
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 23
tgtggtaact ggatagtaaa tcttcc 26
<210> 24
<211> 24bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 24
gctgatgttt aagttttcga agaa 24
<210> 25
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 25
agagcaaaac acaataactc ctctaa 26
<210> 26
<211> 27bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 26
aaatgaagat gtagctagaa gggatac 27
<210> 27
<211> 31bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 27
cagacaagga gagaataact taatattaga g 31
<210> 28
<211> 27bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 28
tcctaaggaa cctatgtgta tattagc 27
<210> 29
<211> 22bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 29
accactacct gcctcaagac ag 22
<210> 30
<211> 22bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 30
acttctctta gcgccttctc ac 22
<210> 31
<211> 22bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 31
gccttacatg ctgaagatgc tc 22
<210> 32
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 32
caaatgatct tcagtttagt accgtg 26
<210> 33
<211> 23bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 33
ttcgtatatt tcacactcca gcc 23
<210> 34
<211> 28bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 34
ttactattaa aactactgtg tcaaccct 28
<210> 35
<211> 23bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 35
aaaatatgga gtctcgggaa aag 23
<210> 36
<211> 24bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 36
aaggaatccc ttcttttgta aaac 24
<210> 37
<211> 21bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 37
tgagtttgag caagttccag g 21
<210> 38
<211> 25bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 38
tgctttcatc tttgaatcag aagac 25
<210> 39
<211> 25bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 39
cagagttcag tgccaaaaat atagg 25
<210> 40
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 40
cagattgcaa cctatcttgt tagaac 26
<210> 41
<211> 21bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 41
atatggatct gggcctcaga g 21
<210> 42
<211> 24bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 42
tctctgcaca cagaggttat atgc 24
<210> 43
<211> 20bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 43
gcaagtgtcc caggcctatg 20
<210> 44
<211> 19bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 44
accgaagtgg gtcctggtt 19
<210> 45
<211> 29bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 45
gacaatatta atccaaatac atcaataag 29
<210> 46
<211> 30bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 46
ctctttactt ttaatcagtt ttaatctttc 30
<210> 47
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 47
agaaacacag agaaagagaa agtgag 26
<210> 48
<211> 28bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 48
cagtggtgtt tactgataga cttagaac 28
<210> 49
<211> 20bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 49
ttcatggcag cctctgctct 20
<210> 50
<211> 21bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 50
acacctgacc atctccgtct t 21
<210> 51
<211> 18bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 51
ctggtcagcg gttgagca 18
<210> 52
<211> 28bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 52
tcatctagta tgtcatcttt tttgagac 28
<210> 53
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 53
gacataactg aagtgactta caagca 26
<210> 54
<211> 22bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 54
ttgccttctc catcaattaa gc 22
<210> 55
<211> 19bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 55
ggactgaaac aggctgggc 19
<210> 56
<211> 26bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 56
gcaaggatta gactgaaaga ttcttc 26
<210> 57
<211> 24bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 57
cgttctgaac cagtaagtat cacc 24
<210> 58
<211> 23bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 58
acccagacag atacaaggga cag 23
<210> 59
<211> 23bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 59
tcataaaatg tgtccagggt cac 23
<210> 60
<211> 18bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 60
tcctgaacac gtggccca 18
<210> 61
<211> 29bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 61
atcttaaatc tttattaacc tattaaggc 29
<210> 62
<211> 22bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 62
tccgtcactt ttctttctca gc 22
<210> 63
<211> 25bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 63
tccactgtta ttttttcact tcttg 25
<210> 64
<211> 27bp
<212> DNA
<213> Artificial sequence
<220>
<223>
<400> 64
cagatacaaa ggaaaattta acaaaac 27
Claims (5)
1. Use of a substance for detecting X in 1) or 2) below:
1) tracing and identifying beef products;
2) identifying a cattle individual;
the X is any one of X1) -X24):
x1) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, and rs 29014143;
x2) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, and rs 29014953;
x3) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953 and rs 17870274;
x4) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, and rs 29021631;
x5) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, and rs 29024289;
x6) is composed of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, and rs 29017633;
x7) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, and rs 41255624;
x8) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624 and rs 29019421;
x9) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421 and rs 29023295;
x10) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295 and rs 29020547;
x11) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547 and rs 29009782;
x12) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, and rs 29020876;
said X13) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, and rs 29018402;
said X14) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, and rs 29025314;
x15) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, and rs 29025593;
x16) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, and rs 29012633;
said X17) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, rs29012633, and rs 29019919957;
x18) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, rs29012633, rs 29019919957, and rs 29010153;
x19) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, rs29012633, rs29019957, rs29010153, and rs 29022322377;
x20) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, rs29012633, rs29019957, rs29010153, rs29022377, and rs 29010275;
said X21) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, rs29012633, rs29019957, rs29010153, rs29022377, rs29010275 and rs 29001970;
x22) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, rs29012633, rs29019957, rs29010153, rs 29022322377, rs29010275, rs29001970, and rs 29009713;
x23) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, rs29012633, rs29019957, rs29010153, rs 29022322377, rs29010275, rs29001970, rs29009713, and rs 29021886;
the X24) consists of rs29019900, rs29010374, rs29016185, rs29010006, rs29010035, rs29023422, rs29014752, rs17871566, rs29014143, rs29014953, rs17870274, rs29021631, rs29024289, rs29017633, rs41255624, rs29019421, rs29023295, rs29020547, rs29009782, rs29020876, rs29018402, rs29025314, rs29025593, rs29012633, rs29019957, rs29010153, rs29022377, rs29010275, rs29001970, rs29009713, rs29021886, and rs 29011378;
the cattle are Simmental cattle, Luxi yellow cattle or Xinjiang black cattle.
2. Use according to claim 1, characterized in that:
the substance for detecting rs29019900 consists of a single-stranded DNA molecule shown in a sequence 1 and a single-stranded DNA molecule shown in a sequence 2;
the substance for detecting rs29010374 consists of a single-stranded DNA molecule shown in sequence 3 and a single-stranded DNA molecule shown in sequence 4;
the substance for detecting rs29016185 consists of a single-stranded DNA molecule shown in a sequence 5 and a single-stranded DNA molecule shown in a sequence 6;
the substance for detecting rs29010006 consists of a single-stranded DNA molecule shown in a sequence 7 and a single-stranded DNA molecule shown in a sequence 8;
the substance for detecting rs29010035 consists of a single-stranded DNA molecule shown in sequence 9 and a single-stranded DNA molecule shown in sequence 10;
the substance for detecting rs29023422 consists of a single-stranded DNA molecule shown in a sequence 11 and a single-stranded DNA molecule shown in a sequence 12;
the substance for detecting rs29014752 consists of a single-stranded DNA molecule shown in sequence 13 and a single-stranded DNA molecule shown in sequence 14;
the substance for detecting rs17871566 consists of a single-stranded DNA molecule shown in sequence 15 and a single-stranded DNA molecule shown in sequence 16;
the substance for detecting rs29014143 consists of a single-stranded DNA molecule shown in a sequence 17 and a single-stranded DNA molecule shown in a sequence 18;
the substance for detecting rs29014953 consists of a single-stranded DNA molecule shown in sequence 19 and a single-stranded DNA molecule shown in sequence 20;
the substance for detecting rs17870274 consists of a single-stranded DNA molecule shown in sequence 21 and a single-stranded DNA molecule shown in sequence 22;
the substance for detecting rs29021631 consists of a single-stranded DNA molecule shown in sequence 23 and a single-stranded DNA molecule shown in sequence 24;
the substance for detecting rs29024289 consists of a single-stranded DNA molecule shown in a sequence 25 and a single-stranded DNA molecule shown in a sequence 26;
the substance for detecting rs29017633 consists of a single-stranded DNA molecule shown in sequence 27 and a single-stranded DNA molecule shown in sequence 28;
the substance for detecting rs41255624 consists of a single-stranded DNA molecule shown in sequence 29 and a single-stranded DNA molecule shown in sequence 30;
the substance for detecting rs29019421 consists of a single-stranded DNA molecule shown as a sequence 31 and a single-stranded DNA molecule shown as a sequence 32;
the substance for detecting rs29023295 consists of a single-stranded DNA molecule shown in a sequence 33 and a single-stranded DNA molecule shown in a sequence 34;
the substance for detecting rs29020547 consists of a single-stranded DNA molecule shown in a sequence 35 and a single-stranded DNA molecule shown in a sequence 36;
the substance for detecting rs29009782 consists of a single-stranded DNA molecule shown as a sequence 37 and a single-stranded DNA molecule shown as a sequence 38;
the substance for detecting rs29020876 consists of a single-stranded DNA molecule shown in sequence 39 and a single-stranded DNA molecule shown in sequence 40;
the substance for detecting rs29018402 consists of a single-stranded DNA molecule shown in a sequence 41 and a single-stranded DNA molecule shown in a sequence 42;
the substance for detecting rs29025314 consists of a single-stranded DNA molecule shown in sequence 43 and a single-stranded DNA molecule shown in sequence 44;
the substance for detecting rs29025593 consists of a single-stranded DNA molecule shown in a sequence 45 and a single-stranded DNA molecule shown in a sequence 46;
the substance for detecting rs29012633 consists of a single-stranded DNA molecule shown in sequence 47 and a single-stranded DNA molecule shown in sequence 48;
the substance for detecting rs29019957 consists of a single-stranded DNA molecule shown in sequence 49 and a single-stranded DNA molecule shown in sequence 50;
the substance for detecting rs29010153 consists of a single-stranded DNA molecule shown in a sequence 51 and a single-stranded DNA molecule shown in a sequence 52;
the substance for detecting rs29022377 consists of a single-stranded DNA molecule shown in sequence 53 and a single-stranded DNA molecule shown in sequence 54;
the substance for detecting rs29010275 consists of a single-stranded DNA molecule shown in sequence 55 and a single-stranded DNA molecule shown in sequence 56;
the substance for detecting rs29001970 consists of a single-stranded DNA molecule shown in a sequence 57 and a single-stranded DNA molecule shown in a sequence 58;
the substance for detecting rs29009713 consists of a single-stranded DNA molecule shown in sequence 59 and a single-stranded DNA molecule shown in sequence 60;
the substance for detecting rs29021886 consists of a single-stranded DNA molecule shown in sequence 61 and a single-stranded DNA molecule shown in sequence 62;
and the substance for detecting rs29011378 consists of a single-stranded DNA molecule shown in sequence 63 and a single-stranded DNA molecule shown in sequence 64.
3. A method for tracing and identifying beef products comprises the following steps: genotyping each bovine to be marketed in a bovine farm with the substance of claim 1 to obtain the genotype of each bovine to be marketed; when the cattle are slaughtered and marketed, taking a meat sample of the cattle, and carrying out genotype identification on the meat sample by adopting the substance to obtain the genotype of the meat sample; if the genotype of the meat sample is the same as that of a certain cattle to be marketed, the meat sample is derived from the cattle;
the cattle are Simmental cattle, Luxi yellow cattle or Xinjiang black cattle.
4. The method of claim 3, wherein: the genotyping is based on a high-throughput Hi-SNP typing platform.
5. A product for beef product traceability identification, which is the substance for detecting X in claim 1;
the cattle are Simmental cattle, Luxi yellow cattle or Xinjiang black cattle.
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Application publication date: 20170926 Assignee: JIANGXI ZODOLABS BIOENGINEERING CO.,LTD. Assignor: INSTITUTE OF QUALITY STANDARD AND TESTING TECHNOLOGY FOR AGRO-PRODUCTS, CHINESE ACADAMY OF AGRICULTURAL SCIENCES Contract record no.: X2022110000089 Denomination of invention: SNP Marker Combinations and Their Applications for Individual and Meat Product Traceability Identification of Beef Cattle Granted publication date: 20210226 License type: Common License Record date: 20221209 |