SNP molecular marker located on porcine No. 7 chromosome and related to total teat number and application of SNP molecular marker
A technology of molecular markers and chromosomes, which is applied in the field of molecular biotechnology and molecular markers, can solve problems such as unsatisfactory results, and achieve the effects of increasing core competitiveness, increasing frequency, and rapid and accurate breeding
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Embodiment 1
[0034] (1) Experimental animals
[0035] The experimental pig group used in the present invention is 2153 purebred plus line Duroc of Wen's Food Group Co., Ltd. Breeding Pig Branch, which is the core group of Breeding Pig Branch.
[0036] In the resource group selected in this experiment, the Canadian Duroc breed pigs, the pigs have free access to food and water, and the entire feeding method and feeding conditions are always consistent, which is a conventional method.
[0037] (2) Sample collection
[0038] The above-mentioned piglet docked tail and ear tissues were collected and soaked in ethanol solution with a volume fraction of 75%, and stored in a -20°C refrigerator for later use.
[0039] (3) 50K SNP typing in the whole pig genome
[0040] The ear tissue or docked tail tissue collected from each individual of the 2153 Duroc breeding pigs selected from the above-mentioned resource groups, the whole genome DNA was extracted by the standard phenol-chloroform method, and ...
Embodiment 2
[0050] Example 2 Target DNA sequence amplification and sequencing
[0051] (1) Primer design
[0052] The DNA sequence of SEQ ID NO: 1 on pig chromosome 7 was downloaded from the Ensembl website (http: / / asia.ensembl.org / index.html). And use primer design software primer premier 6.0 to design primers. The DNA sequences of the designed primers are as follows:
[0053] P001-F: 5'-TTGTGGCGGAGGAGAAAGAGA-3';
[0054] P002-R: 5'-CGGACACCTGAAGTGCTGAT-3';
[0055] (2) PCR amplification
[0056] Add 1 μL of DNA template, 3.4 μL of double distilled water, 5 μL of 2×Tag PCR StanMix with Loading Dye, and 0.3 μL of primers P001-F and P002-R into a 10 μL reaction system. The PCR reaction conditions were as follows: 3 minutes of pre-denaturation at 95°C, 35 cycles of denaturation at 95°C for 30 s, annealing at 60°C for 30 s, extension at 72°C for 30 s, and finally 5 min at 72°C.
[0057] (3) DNA sequence determination
[0058] DNA sequence sequencing identification: carried out in Shen...
Embodiment 3
[0061] Example 3 SNP site g.131G>A effect analysis of molecular markers
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