Method for constructing colorectal cancer susceptibility gene variation library

Abstract

The invention relates to a colorectal cancer susceptibility gene mutation library constructing method. The colorectal cancer susceptibility gene mutation library constructing method comprises the following steps: A, designing primer pairs capable of detecting a mutation region according to the mutation region related to a colorectal cancer; B, calculating to obtain judgment parameters R of the various primer pairs, classifying the primer pairs of which the values of the judgment parameters R are smaller than or equal to 1 into a first group of primer combination liquid, and classifying the primer pairs of which the values of the judgment parameters R are greater than 1 into a second group of primer combination liquid; C, extracting and purifying DNA of a sample to be detected; D, carrying out initial PCR amplification on the purified sample by using the first group of primer combination liquid and the second group of primer combination liquid respectively; E, connecting target fragments via joints to obtain fragments with joints; and F, carrying out library PCR amplification by using mixed liquid of the first group of primer combination liquid and the second group of primer combination liquid to obtain a sequencing library. The library constructed by the colorectal cancer susceptibility gene mutation library constructing method is high in sequencing flux, high in sensitivity and high in specificity, and can be used for detecting free DNA low frequency mutation.

Description

technical field [0001] The invention relates to a method for constructing a colorectal 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 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 throughp...

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Problems solved by technology

[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

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