Multi-omics integration accurate pig breeding method
A technology for piglets, which is applied in the field of multi-omics integrated precision breeding of reproductive efficiency, can solve the problems of incomplete reproductive efficiency of pigs, insufficient understanding and understanding, and achieve the effect of accelerating the breeding process, accelerating genetic progress, and promoting breeding selection
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Embodiment 1
[0083] The acquisition of embodiment 1 SNP molecular marker
[0084] 1. Experimental sample collection
[0085] 375 large white pigs from a pig farm in Shandong were used for SNP typing of HSD17B1 gene. Collect ear tissue samples one by one, put them in 75% alcohol and store them at -20°C for DNA extraction.
[0086] 2. HSD17B1 gene SNP typing
[0087] 2.1 Extraction and detection of pig genomic DNA
[0088] (1) Cut an appropriate amount of pig ear tissue, cut it into pieces and put it into a 1.5mL Axgen tube.
[0089] (2) Take a 50mL BD centrifuge tube, mix proteinase K with a final concentration of 0.4mg / mL and lysis buffer evenly, and add 0.5mL lysis solution to the 1.5mL centrifuge tube containing pig ear tissue for lysis.
[0090] (3) Place the centrifuge tubes in parallel and evenly on the shaking plate of the thermostatic hybridization furnace (seal the tube caps tightly to avoid liquid leakage). Place at 55°C for more than 6 hours (full mixing of the sample during...
Embodiment 2
[0125] Example 2 Detection of HSD17B1 gene expression
[0126] 1. Experimental sample collection
[0127]The large white pigs and Meishan pigs used to detect the expression of HSD17B1 gene came from the pig farm of Beijing Institute of Animal Husbandry and Veterinary Medicine, Chinese Academy of Agricultural Sciences, Wuqing District, Tianjin. The ovaries of sows at 49 days of gestation were collected, all samples were stored in liquid nitrogen, and total RNA was extracted. Three individuals of each breed were collected as biological replicates.
[0128] 2. Detection of HSD17B1 gene expression
[0129] 2.1 Extraction of total RNA from ovarian tissue
[0130] The glassware and tweezers used in the RNA extraction process must be dry-baked at 200°C for 4 hours to inactivate RNase. Eppendorf tubes and pipette tips must use RNase inactivation products. During the entire experiment, experimenters must wear masks and latex gloves.
[0131] (1) Sample collection: Place the collect...
Embodiment 3
[0157] Embodiment 3 detects the kit of relevant SNP site
[0158] The SNP site detection kit of the present invention comprises the following components:
[0159] (1), dNTP (2.5mM);
[0160] (2), 5×PCR buffer;
[0161] (3), FastPfuDNA polymerase (250U);
[0162] (4), SNaPshot mixed solution;
[0163] (5), MgSO 4 Solution (50mM);
[0164] (6), 6×DNA Loading Buffer;
[0165] (7), PCR amplification primer (P 1 and P 2 ): its nucleotide sequence is shown in SEQ ID No.1 and SEQ ID No.2;
[0166] (8), extension primer: its nucleotide sequence is shown in SEQ ID No.3;
[0167] (9), exonuclease (250U);
[0168] (10), shrimp-alkaline phosphatase (100U);
[0169] (11), purification buffer:;
[0170] (12), ddNTP;
[0171] (13), the positive control specimen of the homozygous mutation of the SNP site: a plasmid containing a G base artificially synthesized at the porcine rs323407415 site;
[0172] (14), the positive control specimen of the heterozygous mutation at the SNP sit...
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