Kit for identifying genetype of silky fowl skin color by detecting EDN3 (endothelin 3) gene copy number difference

Abstract

The invention relates to a kit for identifying the genetype of a silky fowl skin color by detecting EDN3 (endothelin 3) gene copy number difference. The kit comprises a reference gene GAPDH (reduced glyceraldehyde-phosphate dehydrogenase) amplification primer (1), a target gene EDN3 amplification primer (2), calibration sample DNA (deoxyribonucleic acid) (3), fluorescent quantitative PCR (Polymerase Chain Reaction) reaction liquid (4) and double distilled water (5). The kit is characterized in that real-time fluorescent quantitative PCR reaction is carried out on a target gene EDN3 and a reference gene GAPDH by using the DNA of a tested sample as a template, Ct values are respectively recorded by using the calibration sample DNA as a reference, the EDN3 gene copy numbers of the tested sample are obtained by calculation, and further the Fm genetype of the silky fowl skin color is judged according to the EDN3 gene copy number difference. The operation of the kit does not need the establishment of a standard curve; the kit is simple, convenient, fast, good in repeatability and high in detection efficiency; and through chicken breed test-crossing testing, the method can accurately reflect the Fm homozygosis and heterozygosis of the skin color of the individual silky fowl. The detection method is conductive to acceleration of screening of homozygosity of the silky fowl skin color, guides target breeding of high-quality silky fowl new variety lines and provides reliable germplasm guarantee for follow-up commercialization mating hybridization.

Description

technical field [0001] The invention relates to a molecular biology kit for detecting the copy number difference of EDN3 gene of black-bone chicken, belonging to the technical field of agricultural biological diagnosis. Background technique [0002] Copy number variation (CNVs) is a newly discovered genomic structural variation that is as important as single nucleotide polymorphisms (SNPs). Human studies have shown that CNVs can lead to single gene diseases and rare diseases with Mendelian inheritance, and at the same time Also associated with many multifactorial diseases and phenotypes such as height. Black skin, black bones, black meat and other black traits are one of the most characteristic appearances of black-bone chickens. This phenomenon of melanin deposition in the body is also called Dermal hyperpigmentation or Fibromelanosis (Fm). In production, effectively distinguishing the purity and purity of skin and black traits will help to speed up the breeding process of...

AI Technical Summary

Problems solved by technology

[0004] Traditional gene copy number detection techniques include western blotting, fluorescent probe hybridization, multiple amplifiable probe hybridization, multiple ligation probe hybridization, etc., but most of them have complex detection process, tedious and time-consuming, high cost, radioactive hazards, false positives, etc. Disadvantages such as positive and low sensitivity

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