A molecular marker related to boar sperm motility and total sperm count and its application
A technology of molecular markers and sperm motility, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc., can solve problems such as the decline of boar semen quality and the decrease of weaned piglets, and achieve high sensitivity and specificity good sex effect
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Embodiment 1
[0032] Example 1: Genotyping Detection
[0033] (1) The total DNA of Duroc boar sperm was extracted using the conventional magnetic bead method sperm genome extraction kit (Wuhan Nanomagnetic Biotechnology Co., Ltd.), the specific steps are as follows
[0034] (1) Take an appropriate amount of semen (5-15 μL) from a Duroc boar into a 1.5 mL centrifuge tube;
[0035] (2) Add 500 μL of lysate (reagents included in the above kit) and 5 μL of proteinase K (20 mg / mL) to the centrifuge tube, shake and mix for 30 seconds, then place in a 65°C oven or metal bath for 30 minutes. ~1h;
[0036] (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;
[0037] (4) Place the No. ① deep-well plate on station 1 of the nucleic acid extractor, and place the deep-well plates containing magnetic beads, washing solution ①, washing solution ②, washing solution ③ and eluent on station 2 respectively. ~6 on. Tu...
Embodiment 2
[0044] Example 2: Application of ALGA0001958 Molecular Marker Typing Method in Association Analysis of Duroc Boar Semen Traits
[0045] (1) Phenotypic pretreatment of semen traits of Duroc boars
[0046] The traits of boar semen belong to repeated measurement traits. It is necessary to use the mixed linear model (MLM). The joint effect of boar station, year and month is used as the fixed effect, the age in days and semen collection interval are used as covariates, and the individual is used as a random effect to calculate the individual random effect. Effects, as newly constructed phenotypes (pseudo-phenotypes) for subsequent genome-wide association analysis (GWAS). The lme4 package in the R statistical environment was used for analysis. The data included 2020 individuals and 105201 semen collection records. The specific model is as follows:
[0047]
[0048] Among them, y ijklm is the original phenotypic value of semen traits (sperm motility and total sperm count) of th...
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