Construction method of lung cancer polygenic variation library

Abstract

The invention relates to a construction method of a lung cancer polygenic variation library, comprising the steps of A, designing a primer pair capable of detecting variation areas of lung cancer related genes according to these variation areas; B, calculating to obtain judging parameters R of the primer pairs, classifying the primer pairs with the judging parameters R less than or equal to 1 as a first group of primer combination liquids, and the primer pairs with the judging parameters R greater than 1 as a second group of primer combination liquids; C, extracting and purifying sample DNA to be detected; D, subjecting the purified sample to initial PCR (polymerase chain reaction) amplification with the first group of primer combination liquids and the second group of primer combination liquids respectively; E, connecting a linker to a target fragment to obtain a fragment with the linker; F, subjecting a mixed liquid of the first group of primer combination liquids and the second group of primer combination liquids to library PCR amplification to obtain a sequencing library. The library constructed by the construction method has high sequencing throughput, high sensitivity and high specificity, and can detect low-frequency variations of free DNA.

Description

technical field [0001] The invention relates to a method for constructing a lung cancer multigene variation library, belonging 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 low cost unma...

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Problems solved by technology

[0008] In order to obtain a template secondary library that can meet the needs of library amplification, the digested amplification products need polyacrylamide gel electrophoresis or agarose gel purification to remove undigested nucleic acids, digested fragments, and nucleases. etc. It can be seen that the current common secondary library preparation methods have the defects of low yield or poor specificity

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