SNPs (Single nucleotide polymorphisms) sites of SPAG6 (Sperm associated antigen 6) gene for screening chicken with excellent sperm quality, heplotype composition thereof and application of heplotype composition
A haplotype combination and genotype technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., can solve problems such as sperm head breakage, sperm structure damage, couple damage, etc.
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Embodiment 1
[0016] Example 1: Identification of SPAG6_SNP1-5 markers closely related to chicken semen quality and primers for detecting genotypes at SPAG6_SNP1-5 sites and compositions containing the primers
[0017] 1. Test data collection
[0018] 390 Beijing oil chicken roosters raised in Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences were selected. All experimental chickens were from the same batch hatched and reared under the same conditions. When the roosters were raised to 43 weeks old, an experimenter collected the semen by abdominal massage, collected it in a semen collection cup, and immediately measured the semen volume, sperm motility, sperm viability and sperm deformity rate.
[0019] 2. Extract the genomic DNA of individuals in the above groups
[0020] Genomic DNA of chicken peripheral blood cells was extracted by high-salt method.
[0021] (1) Digestion Add 30μl chicken anticoagulant blood to a 2ml centrifuge tube...
Embodiment 2
[0033] Example 2: Identification of haplotype combinations of SPAG6_SNP1-5 loci with good semen quality
[0034] 1. Haplotype construction
[0035] Input the genotype data of SPAG6_SNP1-5 loci of all individuals obtained through the above PCR-direct sequencing method into the PHASE program, and the haplotype of each individual can be calculated, and the degree of pairwise linkage disequilibrium between the loci can be calculated at the same time , represented by the normalized linkage disequilibrium coefficient D'.
[0036] The results of pairwise linkage disequilibrium analysis of SPAG6 gene 5 loci at SPAG6_SNP1-5 locus are shown in Table 2. The analysis results show that the linkage disequilibrium coefficient D' between SPAG6_SNP1 and SPAG6_SNP2, SPAG6_SNP4 and SPAG6_SNP5 base sites is 1, which is a complete linkage disequilibrium; the linkage disequilibrium between SPAG6_SNP1 (or SPAG6_SNP2) and SPAG6_SNP3 base sites is in a state of linkage instability. The linkage diseq...
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