rad51l1 gene mutation detection specific primers and liquid phase chip

Abstract

The invention discloses a liquid phase chip and a specific primer for detecting RAD51L1 gene mutation. The liquid phase chip mainly comprises: ASPE primers, microspheres coated by different anti-tag sequences, and an amplification primer, wherein each ASPE primer is composed of a tag sequence at a 5' end and specific primer sequences at a 3' end and aiming at target gene mutation sites, and the specific primer sequences are as follows: SEQ ID NO.9 and SEQ ID NO.10 aiming at a C167T site, SEQ ID NO.11 and SEQ ID NO.12 aiming at an A76T site, SEQ ID NO.13 and SEQ ID NO.14 aiming at a T187G site, and / or SEQ ID NO.15 and SEQ ID NO.16 aiming at a T99G site. The coincidence rate of the detection result of the detection liquid phase chip disclosed by the invention with that of a sequencing method reaches as high as 100%, and parallel detection of a wild type and a mutant 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 RAD51L1 gene mutation detection specific primer and a liquid phase chip. Background technique [0002] RAD51-like 1 (RAD51-like1, RAD51L1), also known as hREC2 or RAD51B, is located on the long arm of chromosome 14 14q23-24, specifically between 68286495 and 69062737 base pairs on chromosome 14, and the RAD51L1 gene encodes the repair double strand An enzyme that breaks DNA, but overexpression of this gene leads to cell cycle G1 delay and apoptosis, suggesting a role for this protein in DNA damage. The protein encoded by RAD51L1 is a member of the Rad51 protein family. RAD51 family members are indispensable conserved proteins for homologous recombination DNA repair, and participate in the homologous recombination repair pathway of double-strand DNA breaks during DNA replication. In addition, the chromosomal variation of RAD51L1 gene is a...

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Problems solved by technology

In addition, the chromosomal variation of RAD51L1 gene is associated with the occurrence of pulmonary chondroid hamartoma and uterine leiomyoma, and RAD51L1 gene polymorphism can increase the risk of breast cancer

[0003] At present, RAD51L1 gene mutation detection methods mainly include: Illumina fiber optic bead chip technology, microarray technology and fluorescence quantitative PCR technology. Although Illumina fiber optic bead chip technology is a high-throughput detection system with high sensitivity and accuracy, it has a low degree of automation. , there are many manual operations, which are difficult to meet the needs of practical applications. Microarray technology has problems such as high cost, complexity, low detection sensitivity, poor repeatability, narrow analysis range, and complex synthesis and fixation of probes. In addition, due to hybridization Located on the surface of the solid phase, with a certain degree of steric hindrance

Fluorescent quantitative PCR technology has the characteristics of high sensitivity, strong specificity, and high degree of automation, but it has the disadvantages of easy sample contamination and high false positive rate, and can only detect one mutation type at a time, which also cannot meet the needs of practical applications.

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