SNP molecular markers related to porcine erythrocyte number traits after polymyocyte infection
A molecular marker, red blood cell technology, applied in the direction of recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as the decline of pig production performance, inability to completely control and eliminate infectious diseases, etc., and achieve high sensitivity and specificity. good effect
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Embodiment 1
[0025] Example 1: Genotyping Detection
[0026] (1) Using phenol extraction method to extract ear tissue DNA from Duroc×Erhualian F2 population
[0027] 1) Grind the ear tissue of Duroc×Erhualian F2 generation pig population (provided by Guangdong Huanong Wens Animal Husbandry Co., Ltd.) in liquid nitrogen, add an equal volume of 1×SET (1mL), proteinase K (10ng / mL) to a final concentration of 200ug / mL, then add 10% sodium dodecyl sulfate (SDS) to a final concentration of 0.5%, and shake well. Incubate the digestion overnight in a 55 °C water bath.
[0028] 2) Add an equal volume of Tris-saturated phenol to the digested tissue sample, slowly invert the centrifuge tube for 15 minutes, centrifuge in a low-temperature refrigerated centrifuge at 4°C and 11,000 rpm for 10 minutes, and carefully transfer the supernatant to another centrifuge tube. Mark the corresponding mark.
[0029] 3) Add an equal volume of phenol / chloroform / isoamyl alcohol (volume ratio 25:24:1), slowly inver...
Embodiment 2
[0039] Example 2: Application of CTTNBP2 molecular marker typing method in correlation analysis of pig immune traits
[0040] (1) Correlation analysis between CTTNB2 molecular marker typing results and immune traits
[0041] The experimental pigs used for the detection and analysis of the association between genotype and immune traits come from the F2 generation population (conventional breed) of the Duroc × Erhualian hybrid bred by Guangdong Huanong Wenshi Animal Husbandry Co., Ltd. The DNA used for genotyping is extracted from the ear sample of the F2 generation of Duroc×Erhualian cross (the “F2 generation of Duroc×Erhualian cross” referred to as “pig” in the text of the manual and the table) is used for routine blood testing Blood for flow cytometric analysis was collected from 35-day-old pigs 4 hours after polymyocyte (PolyI:C) inoculation. Using the method based on the single-marker association regression model, using the gender of the individual as a fixed effect, the G...
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