Method and reagent kit for constructing BRCA 1/2 (breast cancer susceptible genes 1/2) detection libraries

Abstract

The invention discloses a method and a reagent kit for constructing BRCA 1 / 2 (breast cancer susceptible genes 1 / 2) detection libraries. The method includes carrying out fragmentation and tail-end repair on homogenized DNA (deoxyribonucleic acid) samples in DNA fragmentation and tail-end repair systems under the effects of interruption enzymes and tail-end repair enzymes; linking products obtained at the previous step with linker sequences under the effects of DNA ligase; utilizing linker linking products obtained at the previous step as templates and carrying out PCR (polymerase chain reaction) amplification by the aid of primers of linker sequences under the effects of DNA polymerases; hybridizing probe sequences with BRCA 1 / 2 and products obtained at the previous step to trap DNA fragments with the BRCA 1 / 2 and carrying out elution to obtain hybridization trapped products; carrying out cyclization on single strands of the hybridization trapped products under the effects of ligase to obtain the BRCA 1 / 2 detection libraries. The method and the reagent kit have the advantages that steps for constructing the libraries are simple, convenient and speedy, the cost can be effectively reduced, workload can be relieved, and variation types are comprehensive and accurate and are high in flux.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a method and kit for constructing a BRCA1 / 2 gene detection library. Background technique [0002] According to the 2015 cancer statistics released by the National Cancer Center in the "Journal of Cancer for Clinicians", the annual incidence of ovarian cancer in Chinese women is 52,100, and the annual death rate is 22,500, with a mortality rate of about 43%. It ranks first among the three major gynecological tumors . Breast cancer has an annual incidence of 272,000 cases and an annual death rate of 70,700 cases, with a mortality rate of about 26%. In addition, research by the American Cancer Society shows that the 5-year survival rate of patients with type I breast cancer is 88%, and the 5-year survival rate of type IV breast cancer is only 15%, which shows that the higher the grade, the lower the survival rate; The higher the survival rate. Therefore, it is of great sig...

AI Technical Summary

Problems solved by technology

Library construction generally needs to be carried out in steps. After fragmentation by physical fragmentation method and conventional enzyme fragmentation method, it is necessary to further select fragments of appropriate size for screening by gel electrophoresis or magnetic beads, followed by end repair, addition of A, addition of adapters, etc. Steps, many steps and cumbersome, long operation time, easy to cause pollution

[0009] Fragmentation of DNA by physical fragmentation requires investment in a fragmentation instrument, which increases the cost of library construction, instrument maintenance, and manual operation, and the original sample volume required for fragmentation is relatively large

Therefore, DNA fragments must be purified and enriched by magnetic beads before subsequent library construction steps can be performed, which further increases manual operations and fragment loss

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