Construction method of ovarian cancer susceptibility gene mutation library

Abstract

The invention relates to a construction method of an ovarian cancer susceptibility gene mutation library. The construction method comprises the following steps: A, according to mutation regions of related genes of ovarian cancer, designing primer pairs capable of detecting the mutation regions; B, calculating to obtain judgment parameters R of all the primer pairs; classifying the primer pairs with the judgment parameters R smaller than or equal to 1 into a first group of primer combined solution, and classifying the primer pairs with the judgment parameters R more than 1 into a second group of primer combined solution; C, extracting and purifying DNA (Deoxyribonucleic Acid) of a sample to be detected; D, carrying out initial PCR (Polymerase Chain Reaction) amplification on the purified sample through adopting the first group of primer combined solution and the second group of primer combined solution respectively; E, carrying out joint connection on target segments to obtain segments with joints; and F, carrying out library PCR amplification by adopting a mixed solution of the first group of primer combined solution and the second group of primer combined solution to obtain a sequencing library. According to the construction method of the ovarian cancer susceptibility gene mutation library, provided by the invention, the sequencing flux of the constructed library is high, the sensitivity is high and the specificity is high; the constructed library can be used for low-frequency mutation of free DNA.

Description

technical field [0001] The invention relates to a method for constructing an ovarian cancer susceptibility gene variation library, which 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 is aimed at tumor targets and recurrence, metastasis and drug resistance after chemotherapy, while traditional disease diagnosis is based on clinical experience, pathological monitoring , cell detection as the basis, there are limitations of sensitivity, and it is impossible to achieve early detection and early treatment or early prediction and early prevention. [0003] The new generation of high-throughput genetic testing technology, today mainly used Illumina's Hiseq, Miseq series and Life's Ion Torrent TM Represented by the series of sequencers, it has the advantages of high throughput...

AI Technical Summary

Problems solved by technology

[0006] At present, the common methods for detecting ovarian cancer susceptibility genes are Sanger sequencing method and gene chip method. Sanger sequencing method can detect ovarian cancer susceptibility genes regionally, 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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