Method, primer and kit for auxiliary detection of pig backfat thickness character by using pig chromosome 2 gene polymorphic site and application
A polymorphic site, auxiliary detection technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve the effect of reducing breeding costs, improving breeding progress, and low cost
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Embodiment 1
[0038]Example 1. Determination of polymorphic sites, and correlation analysis between polymorphic sites and pig backfat thickness traits 1. Identification of polymorphic sites on pig chromosome 2 (v11.1) g.71588075G>A Sure
[0039] 1. PCR amplification
[0040] According to the upstream and downstream sequence information of porcine chromosome 2 (v11.1) g.71588075G>A in the International Porcine Genome Version 11.1 Reference Sequence, a pair of primers were designed and synthesized as follows:
[0041] U (upstream primer): 5'-GAGATGGCTTCACTGAGG-3' (sequence 1 in the sequence listing);
[0042] D (downstream primer): 5'-ATCAACAAGGCTGAGGAC-3' (SEQ ID NO: 2 in the Sequence Listing).
[0043] Large white pigs (2 heads) were selected as experimental materials. Using the genomic DNA of Large White pig as a template, the upstream primer U and downstream primer D were used for PCR amplification. (Choose the cells of the ear tissue of Large White pigs as a biological sample, take t...
Embodiment 2
[0066] Embodiment 2, establishment of detection method (provide primer pair, kit)
[0067] (1) Establish a detection method using the polymorphic site of the pig chromosome 2 gene to assist in the detection of pig backfat thickness traits, that is, a detection method using the pig chromosome 2 g.71588075G>A in the International Porcine Genome Version 11.1 Reference Sequence A method for detecting backfat thickness traits of pigs with a body weight of 100 kg assisted by polymorphic sites. The detection method is: detecting the deoxyribonucleoside at position 71588075 from the 5′ end of pig chromosome 2 (v11.1) of the pig to be tested Whether the acid is G or A, that is, to detect whether the 71588075th deoxyribonucleotide of the pig chromosome 2 in the reference sequence of the international pig genome version 11.1 of the pig to be tested is G or A from the 5′ end, and determine the gene of the pig to be tested Whether the genotype is GG or AA, the backfat thickness of pigs wit...
Embodiment 3
[0080] Embodiment 3, establishment of breeding method (application of detection method, primer pair, test kit)
[0081] The detection method, the primer pair, and the kit described in the above-mentioned Example 2 are applied to pig breeding to breed pigs with thinner backfat thickness traits.
[0082]The method for cultivating pigs with thinner backfat and thick traits: detect whether the 71588075th deoxyribonucleotide of the pig chromosome 2 (v11.1) of the pig to be tested is G or A from the 5' end, and determine whether the pig to be tested is G or A. Whether the genotype is GG or AA; pigs with AA genotype are selected for breeding, and pigs with thinner backfat and thicker traits can be obtained.
[0083] The method for detecting whether the 71588075th deoxyribonucleotide of the pig chromosome 2 (v11.1) of the pig to be tested is G or A from the 5' end is specifically sequencing analysis; the sequencing analysis includes PCR amplification and Steps for sequencing the PCR ...
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