Method for constructing breast cancer susceptibility gene variation library

Abstract

The invention relates to a breast cancer susceptibility gene variable library construction method. The construction method comprises the following steps that 1, according to variation regions of a breast cancer-related gene, primer pairs capable of detecting the variation regions are designed; 2, judgment parameters R of the primer pairs are calculated, the primer pairs of which values of the judgment parameters are smaller than or equal to 1 are classified into a first primer combination solution, and the primer pairs of which values of the judgment parameters are larger than 1 are classified into a second primer combination solution; 3, a to-be-detected DNA sample is extracted and purified; 4, initial PCR amplification is conducted on the purified sample by adopting the first primer combination solution and the second primer combination solution separately; 5, linker connection is conducted on a target fragment to obtain a fragment with a linker; 6, library PCR amplification is conducted through a mixed solution of the first primer combination solution and the second primer combination solution to obtain a sequencing library. The library constructed through the breast cancer susceptibility gene variable library construction method is high in sequencing flux, sensitivity and specificity and can be used for detecting low-frequency mutation of free DNA.

Description

technical field [0001] The invention relates to a method for constructing a breast cancer susceptibility gene variation library, and belongs to the field of biotechnology. Background technique [0002] At present, the morbidity and mortality of tumors are higher than those of other diseases. The development of non-invasive diagnostic strategies for tumor gene diagnosis and development aims at tumor targets and recurrence, metastasis and drug resistance after chemotherapy, while traditional disease diagnosis is based on clinical experience and pathological monitoring. , Cell detection as the basis, there are sensitivity limitations, and it is impossible to achieve early detection and early treatment or early prediction and early prevention. [0003] A new generation of high-throughput gene detection technology, currently mainly based on Illumina's Hiseq, Miseq series and Life's Ion Torrent TM Represented by a series of sequencers, it has the advantages of high throughput and...

AI Technical Summary

Problems solved by technology

[0006] At present, the common methods for detecting breast cancer susceptibility genes are Sanger sequencing method and gene chip method. Sanger sequencing method can detect the region of breast cancer susceptibility genes, but Sanger sequencing method can only detect a certain region of a sample at a time. Sequencing, so the detection efficiency is low and the cost is high

Moreover, due to the limitations of the sanger sequencing principle, double peaks or even multiple peaks will appear when detecting the signal of a sample with a heterozygote, making the sample unable to be accurately identified, resulting in low detection sensitivity (20%)

Gene chip method, this method can only design detection probes for one or several known mutation sites to form a detection chip, so it cannot be used to detect unknown mutation sites, and the detection results are not comprehensive and accurate enough

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