ATR (ataxia telangiectasia-mutated and Rad3-related) gene mutation detection specific primers and liquid chip

Abstract

The invention discloses an ATR (ataxia telangiectasia-mutated and Rad3-related) gene mutation detection liquid chip and specific primers. The liquid chip mainly comprises each ASPE (allele specific primer extension) primer formed by tag sequences at 5' terminal and specific primer sequences aiming at target gene mutation sites at 3' terminal, microspheres coated by anti-tag sequences as well as amplification primers, wherein the specific primer sequences are SEQ ID NO.7 and SEQ ID NO.8 aiming at G632A sites, SEQ ID NO.9 and SEQ ID NO.10 aiming at T7301G sites and / or SEQ ID NO.11 and SEQ ID NO.12 aiming at G946A sites. The liquid chip has the advantages that the coincidence rate of the detection results of the detection liquid chip provided by the invention and a sequencing method is as high as 100%; and parallel detection of wild types and mutant types of a plurality of mutation sites is achieved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for ATR gene mutation detection and a liquid phase chip. Background technique [0002] The ATR gene is located on chromosome 3 (3q22-q24), responsible for encoding ataxia telangiectasia and related protein Rad3 (Ataxiatelangiectasia-mutatedandRad3-related, ATR), ATR protein belongs to phosphatidylinositol 3-kinase-related kinase (phosphatidylinositol3 -kinaserelatedkinase, PIKK) protein family member, is the main member of the DNA damage checkpoint, they are activated by different types of DNA damage, by phosphorylating the corresponding downstream proteins Chk1 and Chk2, etc., regulate each checkpoint of the cell cycle, causing the cell cycle Therefore, ATR plays a crucial role in maintaining the stability of the genome. At present, studies have shown that ATR gene mutations are related to the occurrence and development o...

AI Technical Summary

Problems solved by technology

[0003] At present, the detection methods of ATR gene mutation mainly include: PCR-RFLP, direct sequencing method and fluorescent quantitative PCR technology. PCR-RFLP method is based on the change of restriction enzyme recognition site caused by gene mutation, such as site loss or generation New site, a specific fragment is amplified by PCR, and then the amplified product is digested with restriction endonuclease, and the size of the fragment is observed by electrophoresis. Genotype, but this method cannot be used for the detection of gene mutations that do not generate new restriction sites

However, other PCR-based detection technologies, such as direct sequencing and fluorescent quantitative PCR technology, have the disadvantages of low sensitivity, easy contamination of samples, and high false positive rate.

Again, the above methods all have limitations in detection throughput, and can only detect one mutation type at a time, which cannot meet the needs of practical applications.

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