A Molecular Marker Related to the Traits of Total Teat Number in Pigs and Its Application
A technology of molecular markers and nipples, applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc., to achieve high sensitivity and good specificity
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Embodiment 1
[0029] Example 1: Genotyping Detection
[0030] (1) Using the magnetic bead method genome extraction kit to automatically extract the total DNA from tissue samples of American Large White pigs (from the experimental pig farm of Huazhong Agricultural University), the specific steps are as follows:
[0031] (1) Take an appropriate amount of tissue samples into a 1.5mL centrifuge tube;
[0032] (2) Add 500 μL of lysate and 5 μL of proteinase K (20 mg / mL) to the centrifuge tube, shake and mix for 30 seconds, place in a 65°C oven or metal bath, and lyse for 30 minutes to 1 hour;
[0033] (3) After the lysis is completed, transfer all the supernatant to a deep-well plate (marked as ①), and add 350 μL of isopropanol to each well;
[0034] (4) Put the No. ① deep-well plate on station 1 of the nucleic acid extractor, and place the deep-well plates with magnetic beads, washing solution ①, washing solutions ② and ③ and eluent on station 2~ 6 on. Turn on the power of the instrument, an...
Embodiment 2
[0042] Example 2: Application of the H3GA0022648 molecular marker typing method in the association analysis of the total teat number traits of American Large White pigs
[0043] (1) Phenotypic pretreatment of traits of total teats in American Large White
[0044] The total teat traits of American Large White pigs need to use the mixed linear model (Mixed Linear Model, MLM), the specific steps are: use the joint effect of field, birth year and birth season, gender, and birth order as fixed effects, and individual effects as random Effect, the estimated breeding values (Estimated breeding values, EBV) of individuals with phenotype records estimated by the model are added to the corresponding estimated residuals (residuals), as the corrected phenotype values (Corrected phenotypes,) of these individuals, using for subsequent GWAS analysis. The DMUAI module under the DMU statistical environment was used for Restricted Maximum Likelihood estimation (Restricted Maximum Likelihoo...
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