A primer, method, kit and application for enriching target regions of brca1 and brca2 genes

Abstract

The invention discloses a primer, a method and a kit for enriching BRCA gene target regions and applications thereof, and relates to the technical field of biology. The primer has sequences shown as SEQ ID NO.1 to SEQ ID NO.52, and refers to five groups of primer pairs obtained by optimally combining 26 pairs of primers in one set of which amplification segments include exons and intron regions on side wings of the exons of BRCA1 and BRCA2 genes. Through the five primer groups, the PCR (Polymerase Chain Reaction) number is reduced to five; during amplification of a sample to be tested, the amplification segments of five groups of amplification products obtained through multiple PCR amplification of each group of primers in the five groups of primers are remarkably different from one another, so that the quality of the amplification products is easy to monitor, and 100-percent enriching of BRCA1 and BRCA2 gene target regions in the sample to be tested is realized; gene sequence mutation can be detected efficiently and easily with high accuracy and low cost.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer, method, kit and application for enriching BRCA1 and BRCA2 gene regions by multiplex PCR. Background technique [0002] BRCA1 and BRCA2 are important tumor suppressor genes involved in the process of chromosome damage repair. Mutations in BRCA1 and BRCA2 may result in the reduction or loss of gene function, thereby increasing the risk of cell cancer. Existing data show that the germline mutations of these two genes are the most important factors causing hereditary breast and ovarian cancers, and mutation carriers have a life-time chance of developing breast cancer as high as 87% and ovarian cancer as high as 44% . Therefore, detection of BRCA1 / 2 gene mutations in high-risk groups has important guiding significance for cancer prevention, discovery and treatment. [0003] Most of the mutations found in BRCA1 and BRCA2 are single base mutations or deletions and insertions of...

AI Technical Summary

Problems solved by technology

In this method, each pair of primers is amplified separately, and the number of PCR reaction tubes required for each sample is large, which is not conducive to large-scale operations, because the amplicons overlap, and the setting of the amplification length determines the number of multiple amplification products. Individual fragments cannot be analyzed by simple methods such as agarose gel electrophoresis, so it is not suitable for multiplex PCR to reduce the number of reaction tubes

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