Rapid detection kit for identifying SNP characteristic sites of Y chromosome haplotype pedigree and method

Abstract

The invention discloses a kit for rapidly detecting the SNP site of a Y chromosome and a method, and the method is applied to the rapid identification of Y chromosome haplotype pedigrees of male individuals in forensic science. The rapid detection kit for the identification of the characteristic sites of the Y chromosome haplotype pedigrees of East Asian males includes primers corresponding to theSNP characteristic sites used for amplification detection of the Y chromosomes and probes of two mutation type sequence characteristics representing the SNP sites. The using method includes adding asample into plates having 96 or 384 pores, and performing an amplification reaction in a PCR instrument after a PCR reaction system is well mixed; putting the plates into the quantitative PCR instrument having an SDS scanning function to read a fluorescence signal after the amplification is finished; and automatically analyzing the Y-SNP haplotype of the sample by the instrument according to the fluorescence signal, and outputting a result in a spreadsheet. Through the kit, the detection can be completed by one step amplification, purification and template preparation are not needed, and the detection process of the Y-SNP can be simplified.

Description

technical field [0001] The invention belongs to the technical field of detection, and in particular relates to a kit and a method for rapidly detecting SNP characteristic sites for identifying Y chromosome haplotype pedigree. Background technique [0002] At present, there are several methods for realizing individual pedigree identification and detection of Y chromosome and mitochondrial SNP: [0003] 1. Sequencing method (generation sequencing): Perform PCR amplification on the target site segment, then use a purification kit to purify the PCR product to remove various components in the reaction system, and then use sequencing primers and sequencing reagents based on the PCR product as a template The sequencing reaction was carried out with the cartridge, and the sequencing reaction product was purified by the purification kit, then mixed with deionized formamide and denatured, and then sequenced on the sequencer. This technology involves many steps, it is difficult to ac...

AI Technical Summary

Problems solved by technology

This technology involves many steps, it is difficult to achieve large-scale high-throughput rapid detection, and the cost is also high

[0004] 2. Sequencing method (next-generation sequencing): NGS can realize high-throughput detection of large samples, but this technology requires a very complicated process of library construction and sequencing, and ultimately generates a large amount of data. Data analysis must be completed by professionals

The technology is currently expensive and difficult to promote and use on a large scale

[0005] 3. SNP chip detection method: chip technology is to design the site information that needs to be detected as a probe, and gather the information of multiple sites on one chip to achieve large-scale rapid detection. However, chip hybridization, data reading and data interpretation All require very professional technicians to complete independently

First, Snapshot technology requires two-step PCR amplification (one-step detection product amplification and one-step extension reaction amplification), and two-step enzymatic purification (one-step detection amplification product purification and one-step extension product purification); secondly, multiplex The amplified system needs to adjust the ratio of primers and templates according to the situation so that the final detection signal intensity remains consistent; third, the Snapshot detection results need to manually check the reliability of the original data in order to have a guarantee for data quality; Fourth, in the process of multiple amplification, some site information is often missing, and time-consuming and laborious repeated experiments are required to supplement a large number of missing site information

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