SV40 molecular marker in NELFA (negative elongation factor complex member A) gene related to litter size of mini pigs and application
A technology of molecular markers and litter size, applied in the direction of recombinant DNA technology, DNA/RNA fragments, microbial determination/inspection, etc., can solve the problems of slow growth, slow development of improved pig breeds, pig breed improvement and breeding start Late and other problems, to achieve the effect of easy operation
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Embodiment 1
[0032] Embodiment 1 is the detection method of the present invention, and the steps are as follows:
[0033] 1. Genomic DNA extraction from pig ear tissue
[0034] Genomic DNA from pig ear tissue was extracted using Tiangen Spin Column Tissue Genomic DNA Extraction Kit (catalogue number: DP304), and the steps were as follows:
[0035] (1) Mince an appropriate amount of pig ear tissue and place it in a 1.5mL centrifuge tube, add an appropriate amount of buffer GA (50mg ear tissue with 200μL buffer GA), shake thoroughly to suspend the tissue fragments;
[0036] (2) Add 4 μL RNaseA (100 mg / mL), shake for 15 s, and place at room temperature for 5 min to remove RNA in the tissue;
[0037] (3) Add 20 μL of Proteinase K solution, mix well and place in a 56°C water bath for 1 to 3 hours until the tissue is completely dissolved, then briefly centrifuge to remove water droplets on the tube wall;
[0038] (4) Add 200 μL cell lysate GB, mix well by inverting, and bathe in 70°C water bat...
Embodiment 2
[0055] Example 2 Detection and Application of Structural Variation SV40
[0056] According to the method of Example 1, with the porcine genomic region structural variation site (chr8: 1111424-1111463) as the candidate site, SV40 was detected by PCR technology in the population (226 fragrant pigs, 106 individuals with high litter size, Structural variation in individuals with low litter size (120 pigs), blood samples from 226 Xiang pigs were collected from Zijiang County. Using the SV40 candidate region, applying the present invention to establish the detection technology to analyze the distribution frequency of the structural variation SV40 in the population, the results show that the genotyping results show that there are 3 genotypes in the high-yield population and low-yield population, and all of them are mainly type II Genotype (Table 1).
[0057] Statistics of the litter size of the first three litters of Xiang pigs and calculation of the average (Table 2), found that am...
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