Breeding method for sifting tight duck by detecting saltant duck gene
A mutant and genotyping technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems that have not yet been reported on Adiponectin gene single nucleotide research, to achieve fat content reduction, easy genotyping, The effect of simple detection methods
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Embodiment 1
[0041] Step 1: Design of PCR primers
[0042] According to the complete sequence of the duck Adiponectin genome cloned in our laboratory, a pair of primers were designed as follows:
[0043] Primer A: DuadpF 5′-ACGAGCAGAACCACTACGAC-3′
[0044] DuadpR 5′-CTGAGGTGCAGCAAGACAGA-3′
[0045] The second step: the extraction of duck genomic DNA, take 20 μl fresh blood, add ACD anticoagulant anticoagulant, add 600 μl poultry lysate, described poultry lysate is pH8.0 10mM Tris·Cl, pH8.0 0.1M EDTA and Add proteinase K to the mixed solution with a concentration of 0.5% (m / v) SDS to a final concentration of 100-200 μg / ml, mix well and digest at 55°C for 6-10 hours until there are no sticky lumps in the solution, then cool the solution Bring to room temperature, add an equal volume of phenol, invert the centrifuge tube repeatedly until the two phases mix to form an emulsion, centrifuge at 12,000 rpm for 10 min at room temperature. Take the supernatant, and then use an equal vol...
Embodiment 2
[0080] 1. The duck Adiponectin gene single nucleotide polymorphism was discovered by using the above method.
[0081] Using primer A for PCR-SSCP analysis, we found 6 genotypes ( figure 1 ). It was found that there were 5 nucleotide mutations in this fragment, namely G→A (position 430), A→G (position 457), C→T (position 507), T→C (position 523), T→ C (position 540), these mutations lead to polymorphisms in the Adiponectin coding region. Sequencing results found that BB was identical to the sequence uploaded by GenBank, which was defined as wild type, and the others were defined as mutant ( figure 2 ).
[0082] 2. Discovered the relationship between the single nucleotide polymorphism of Adiponectin gene and production performance
[0083] We performed least squares analysis on the relationships between the found Adiponectin SNPs and traits such as live weight, carcass weight, half-eviscerated weight, liver weight, sebum weight, sebum percentage, abdominal fat weight, and a...
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