Mitochondrial SNP fluorescence labeling composite amplification kit and application thereof

Abstract

The invention relates to a mitochondrial SNP fluorescence labeling composite amplification kit capable of simultaneously detecting 35 SNP sites and main application thereof. The kit employs 35 pairs of specific primers for genetic typing of human mitochondrial SNP sites. The kit comprises a PCR amplification reaction reagent, FAM color composite amplification primers, HEX color composite amplification primers, a positive control, a negative control and fluorescent internal standard. An application method of the kit comprises the following steps: carrying out PCR composite amplification on mitochondrial DNAs extracted from a variety of to-be-detected materials; and collecting fluorescence signals through electrophoresis by using a genetic analyzer, which enables accurate allele typing of each site to be realized. Compared with a conventional mitochondrial SNP sequencing method, the kit provided by the invention is more simple, convenient and economic, uses less templates and has a higher success rate. As a mitochondrial detection system, the kit carries out simultaneous amplification in two tubes and simultaneous electrophoresis, diversity of haplotype reaches 99.64%, and the kit can be used as a kit for identification of a same maternal line following an autosomal STR and a Y chromosome STR.

Description

technical field [0001] The present invention belongs to the technical field of biological in vitro diagnosis, embodies the combined application of the third-generation genetic marker SNP (single nucleotide polymorphism, single nucleotide polymorphism) and fluorescence detection technology, and mainly relates to a method for identifying the maternal lineage of mitochondrial DNA SNP fluorescent labeling compound amplification system. Background technique [0002] At present, the kits used for individual identification in forensic science are all based on STR, but as the third generation of genetic markers, single nucleotide polymorphism (single nucleotide polymorphism, SNP) has a larger number and lower mutation rate than STR. Low, genetically stable, easy to automate, suitable for the analysis of highly degraded samples, etc., its advantages in forensic medical examination are becoming more and more obvious, such as inferring the ethnic origin of samples, identifying physical...

AI Technical Summary

Problems solved by technology

[0005] Therefore, in order to improve the identification ability, the applicant conducted in-depth research on the genetic polymorphisms of human mitochondria in various regions of the country, and selected 35 SNP sites with higher polymorphic sites to form a PCR multiplex amplification system. And based on this, the mitochondrial SNP fluorescent labeling composite amplification kit was developed. This kit has not been reported and used in the world at present, so it is a problem that people in the field have been expecting to solve, and has great application prospects.

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