Detection method and primers for evaluating enrichment result of porcine histone modification

Abstract

The invention provides a detection method and primers for evaluating an enrichment result of porcine histone modification. The method comprises the following steps: enriching DNA combined with targetprotein by using a pig tissue sample through a ChIP-seq method, carrying out quantitative detection by adopting a fluorescent quantitative QPCR method, and judging the result, wherein a calculation method of the quantitative detection result adopts a delta delta Ct method for judgment, and the result is judged as follows: if the effective precipitation enrichment multiple of positive primers and negative primers is greater than or equal to 2, the enrichment effect is considered to reach the standard, and if the effective precipitation enrichment multiple of positive primers and negative primers is less than 2, the enrichment effect is considered to not reach the standard. According to the method, the v histone modification level can be specifically detected, and the DNA region enrichment effect can be effectively judged, so that important reference is provided for ChIP-seq, the time and economic cost caused by poor sequencing result due to poor enrichment effect are greatly saved, andan effective way is provided for developing pig epigenomics research.

Description

technical field [0001] The invention relates to the technical field of ChIP-seq, in particular to a detection method and primers for evaluating the enrichment result of porcine histone modification. Background technique [0002] Fat deposition and muscle growth are one of the most important economic traits of pigs, which directly affect the quality of pork and the level of its economic value. Therefore, it is a very important subject to study the genetic traits of pig fat and muscle tissue through genetics. [0003] Among them, epigenetics research is a newly developed scientific field after genetics, which is to study the heritable expression of genes without changing the nucleotide sequence of genes. Histone modification is an important regulatory mode of epigenetic regulation, which can regulate gene expression through methylation and acetylation modification. H3K4ME3 and H3K27ac are important epigenetic modifications of histone modification, which have been expressed i...

AI Technical Summary

Problems solved by technology

[0005] However, the main materials of ChIP technology research are concentrated on cell or tissue samples of model animals, and the processing methods are relatively mature, but the application in porcine muscle and adipose tissue is less, especially adipose tissue has high requirements for pre-treatment and is difficult to break. It is difficult to extract complete nuclei and the stability between materials is not very good. These factors restrict the application of this technology in porcine tissues

In particular, the cost of ChIP-seq is relatively high. If effective quality control is not done before, it will bring great losses to see the success or failure of the experiment through the sequencing results after large-scale sample enrichment

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