Molecular marker related to white hair phenotypes of buffaloes and application
A molecular marker, buffalo technology, used in animal breeding, genetics and molecular biology to achieve the effect of accurate methods
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Embodiment 1
[0030] Example 1 Obtaining Molecular Markers Related to Buffalo White Coat Color Phenotype
[0031] 1. Sample collection
[0032] In the buffalo producing area in southern China, 63 swamp-type buffaloes were randomly collected, including 41 tile-gray buffaloes and 22 white buffaloes. The sample is the ear epithelial tissue, which is placed in a sample tube with 75% alcohol and stored at -80°C.
[0033] 2. Buffalo Genomic DNA Extraction
[0034] (1) Solution preparation: tissue lysate (1L): 1.2114g solid Tris-Base (121.1), 37.22g solid disodium ethylenediaminetetraacetic acid (EDTA, 372.24), 5.0g solid sodium dodecyl sulfate (SDS ) was dissolved in 800mL of double distilled water, after complete dissolution, additional double distilled water was added to make the volume to 1L, and the pH was adjusted to 8.0.
[0035] Proteinase K solution (20mg / mL): 100mg protein K yellow crystals, dissolved in 5mL deionized water, aliquoted and stored at -20°C.
[0036] (2) Take 200 mg of ...
Embodiment 2
[0062] Example 2 Verification of SNP marker g.9953331
[0063] The g.9953331 site genotyping results are shown in Table 1.
[0064] Table 1 SNP site g.19953331 genotype frequency distribution
[0065]
[0066] *Note: C-mutant, T-normal.
[0067] According to the statistical results of the data of the present invention, when the g.19953331 marker genotype is CC, there is 100% confidence that the individual can be inferred to be white coat color, and when the g.19953331 marker genotype is CT, there are 95.2% (20 / 21×100% =95.2%), it can be inferred that the individual has a white coat color, and when the g.19953331 marker genotype is TT, it can be inferred that the individual is a tile-gray coat color with 100% confidence.
Embodiment 3
[0068] Embodiment 3 Verification of SNP marker g.19970628
[0069] The g.19970628 site genotyping results are shown in Table 2.
[0070] Table 2 SNP site g.19970628 genotype frequency distribution
[0071]
[0072] *Note: A-mutant, G-normal.
[0073] According to the statistical results of the data of the present invention, when g.19970628 marker genotype is AA, there is 100% certainty that the individual can be inferred to be white coat color; when g.19970628 marker genotype is AG, there are 95.2% (20 / 21×100% =95.2%), it can be inferred that the individual has a white coat color, and when the g.19970628 marker genotype is GG, it can be inferred that the individual is a tile-gray coat color with 100% confidence.
[0074] Therefore, the two marker genotypes can be detected in the Chinese buffalo population, and the retention of white homozygous buffalo individuals can rapidly increase the proportion of homozygous white coat color genes.
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