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Method of using digital PCR (polymerase chain reaction) to identify single-copy gene mutation genotypes

A single-copy gene, single-copy technology, applied in the field of microdroplet digital PCR, can solve the problems of low frequency and low adaptability

Inactive Publication Date: 2018-08-10
TARGETINGONE TECH (BEIJING) CORP
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  • Abstract
  • Description
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Problems solved by technology

Due to the majority of mutations, the fitness of the individual is very low, so the frequency is very low due to the effect of negative selection

Method used

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  • Method of using digital PCR (polymerase chain reaction) to identify single-copy gene mutation genotypes
  • Method of using digital PCR (polymerase chain reaction) to identify single-copy gene mutation genotypes
  • Method of using digital PCR (polymerase chain reaction) to identify single-copy gene mutation genotypes

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Embodiment Construction

[0032] In order to enable those skilled in the art to better understand the technical solutions in the application, the present invention will be further described below in conjunction with the embodiments. Obviously, the described embodiments are only a part of the embodiments of the application, rather than all embodiments . Based on the embodiments in this application, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts shall fall within the scope of protection of this application. The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0033] Example Identification of Human Leukocyte Antigen HLA-B27 Genotype by Digital PCR

[0034] An example of micro-droplet digital PCR reaction conditions for testing the performance of primers and probes is as follows: first, extract nucleic acid from clinical samples with a nucleic acid extraction kit, and then prepare a PC...

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Abstract

The invention provides a method of using digital PCR (polymerase chain reaction) to identify single-copy gene mutation genotypes, comprising: designing a digital PCR amplification primer and a probe for a sample's single-copy gene mutation site to be detected, designing a digital PCR amplification primer and a probe for the sample's single-copy housekeeping gene, and subjecting the mutation site and the housekeeping gene to dual-channel double digital PCR; determining whether the sample to be detected has the single-copy gene mutation site according to presence or absence of digital PCR amplification of the single-copy gene mutation site; if yes, identifying the genotype of the single-copy gene to be detected by comparing the copy number of the single-copy gene to be detected to the copy number of the single-copy housekeeping gene. The method has the advantages of good accuracy, good convenience, high speed, low cost and the like and is suitable for genetic disease screening and prevention and genetic diagnosis.

Description

technical field [0001] The invention relates to the technical field of micro-droplet digital PCR, in particular to a method for identifying single-copy gene mutation genotypes by using digital PCR. Background technique [0002] The genome (Genome) is generally defined as the entire base sequence of all chromosomes in a haploid cell, that is, the haploid genome. A single-copy gene refers to a gene that has only one copy in the haploid genome, and the single-copy gene accounts for 50-80% of the genome. For example, in the human haploid genome, about 60-65% of the gene sequences belong to a single Copy genes, such as most gene sequences that code for proteins, are single-copy genes. For example, human leukocyte antigen HLA gene, hereditary deafness gene GJB2, etc. A huge amount of genetic information is stored in a single copy of a gene, encoding a variety of proteins with different functions. Corresponding to single-copy genes are multi-copy genes, that is, genes with multi...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858C12Q1/6851C12Q1/6881
CPCC12Q1/6851C12Q1/6858C12Q1/6881C12Q2531/113C12Q2537/143C12Q2537/16C12Q2563/159
Inventor 祝令香朱宏艳杨文军王勇斗
Owner TARGETINGONE TECH (BEIJING) CORP
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