A SNP marker related to the farrowing traits of Erhualian sows and its detection method and application
A Erhualian sow and marker technology, which is applied in the field of molecular biology, can solve the problems of not being able to determine the main effect gene and its key mutation sites, and achieve important economic benefits and social value
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Embodiment 1
[0031] 1. Source of experimental animals
[0032] Jiaoxi Erhualian pig professional cooperative in Changzhou, Jiangsu.
[0033] Calculate the breeding value of 177 Erhualian sows, the calculation model is
[0034] Y (total litter size) = parity (parity) + farm (field) + year (annual) + season (season) + age (sow farrowing age) + sire (mating boar) + permanent effect (female Pig permanent effect) + additive effect (individual additive effect) + e (residual),
[0035] These included fixed effects - parity, farm / year / season sows farrowed, covariates - age of sows farrowed, random effects - boar with mating, permanent effects - sows, individual additive genetic values. Individuals with the top 10% and bottom 10% of breeding values were selected. Genomic DNA extraction
[0036] Ear tissue samples from 36 sows were collected, placed in centrifuge tubes filled with 70% alcohol, and stored in a -20°C refrigerator for later use.
[0037] Using the traditional phenol / chloroform m...
Embodiment 2
[0064] This example is the verification of the SNP site g.37502410A / C obtained in Example 1 in the population of Erhualian sows.
[0065] 1. Extraction of Genomic DNA from Erhualian sows
[0066] Ear tissue samples from 197 purebred Erhualian sows with accurate records of total litter size were collected, placed in centrifuge tubes filled with 70% alcohol, and stored in a -20°C refrigerator for later use. Genomic DNA from ear tissue was extracted by the above method, and after quality and concentration detection, the concentration was diluted to 30 ng / μL and stored at -20°C for future use.
[0067] 2. PCR amplification and sequencing of the target fragment
[0068] Using the extracted DNA as a template, carry out PCR amplification according to the designed primers: take 2.5 μL of DNA template, 1.25 μL of each primer shown in SEQ ID NO: 2 and SEQ ID NO: 3, 25 μL of PCR Mix reagent, double distilled 20 μL of water; set up the PCR amplification system: pre-denaturation at 96°C ...
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