Screening method and application of meat duck real-time fluorescent quantitative PCR reference genes

Abstract

The invention belongs to the technical field of animal gene engineering, and discloses a screening method and application of meat duck real-time fluorescent quantitative PCR reference genes. The method comprises the following steps of selecting eight candidate reference genes by utilizing a meat duck transcriptome database, designing a real-time fluorescent quantitative PCR specific primer by taking the eight candidate reference gene sequences as templates, performing a fluorescent quantitative PCR experiment, and finally performing data analysis by applying geNorm, NormFinder and BestKeeper software to screen out the most stable reference genes, namely HMBS and YWHAZ. According to the method disclosed by the invention, a screening system of the stable reference genes of the meat ducks is established, and the stable reference genes are provided for subsequent research on meat duck gene expression by adopting fluorescent quantitative PCR.

Description

technical field [0001] The invention relates to the technical field of animal genetic engineering, in particular to a screening method and application of real-time fluorescent quantitative PCR internal reference genes of meat ducks. Background technique [0002] In animal genetics and breeding, the exploration of gene expression patterns is common and important. Real-time fluorescent quantitative PCR is the most commonly used nucleic acid quantitative technology in the study of gene expression patterns, which has higher sensitivity, better specificity and wider detection range. Some possible differences in initial sample volume, RNA integrity, and cDNA synthesis efficiency will affect the accuracy of real-time fluorescent quantitative PCR results. Therefore, screening suitable internal reference genes is an important prerequisite for real-time fluorescent quantitative PCR. [0003] Many studies have screened some reference genes, such as actin (β-actin), ribosomal RNA (18S...

AI Technical Summary

Problems solved by technology

However, a large number of studies have shown that there is no internal reference gene that can satisfy constant expression under all experimental factors, so choosing the wrong internal reference gene not only fails to correct the key genes, but also leads to inaccurate test results

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