SNP molecular marker related to number of healthy piglets and live piglet rate and application thereof
A molecular marker, rate-related technology, applied in recombinant DNA technology, microbial determination/inspection, biochemical equipment and methods, etc. The effect of increasing frequency and improving reproductive capacity
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Embodiment 1
[0028] (1) Experimental animals
[0029] The experimental pig group used in the present invention is 413 purebred Duroc pigs from the Breeding Pig Branch of Wen's Food Group Co., Ltd., which is the core group of the Breeding Pig Branch.
[0030] In this experiment, the Duroc pigs in this resource group were selected, and the pigs had free access to food and water, and the entire feeding method and feeding conditions were always consistent, which was a conventional method.
[0031] (2) Sample collection
[0032] 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.
[0033] (3) 50K SNP typing in the whole pig genome
[0034] For each of the 413 Duroc pigs in the above-mentioned resource groups, ear tissues or tail tissues were collected, and the whole genome DNA was extracted by the standard phenol-chloroform method, and the DNA content of each sampl...
Embodiment 2
[0044] Example 2 Target DNA sequence amplification and sequencing
[0045] (1) Primer design
[0046] The DNA sequence of SEQ ID NO: 1 on pig chromosome 15 was downloaded from the Ensembl website (http: / / asia.ensembl.org / index.html), and primers were designed using the primer design software primer premier 6.0. The DNA sequences of the designed primers are as follows:
[0047] P001-F: 5'-CCTGTGGTAGACCATGCTCCAA-3',
[0048] P002-R: 5'-GGGACACAGGACAAAGGGATG-3';
[0049] (2) PCR amplification
[0050] 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: 5 minutes of pre-denaturation at 94°C, 35 cycles of denaturation at 94°C for 30 s, annealing at 58°C for 30 s, extension at 72°C for 45 s, and finally 5 min at 72°C.
[0051] (3) DNA sequence determination
[0052] DNA sequence sequencing identification: carried...
Embodiment 3
[0055] Example 3 SNP site g.118G>A effect analysis of molecular markers
[0056] According to Table 1, the phenotypic effect of SNP site g.118G>A dominant allele type AA can increase the number of healthy piglets by 3.07 and increase the live piglet rate by 20%. If the profit per piglet is 200 yuan, then Only aiming at the number of healthy piglets can increase the profit of more than 600 yuan per sow, and for large-scale pig farms with 10,000 sows, the economic benefit can be increased by more than 6 million yuan. Therefore, through molecular marker-assisted selection or genomic selection, gradually selecting and retaining pigs with genotype AA in the population can significantly increase the allele frequency of allele A, and increase the number of healthy piglets and live piglets in sows. The rate can speed up the progress of pig improvement and thus effectively improve the economic benefits of pig breeding.
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