A capture probe for detecting gene mutation by high-throughput sequencing and its application

Abstract

The invention relates to the technical field of gene detection, in particular to a capture probe for high-flux gene mutation sequencing and detecting and application thereof. The capture probe is a single-chain probe body mixture which comprises probe bodies for a positive-sense strand and probe bodies for a negative-sense strand, wherein the probe bodies are sequentially designed according to thenucleotide sequences for the positive-sense strand and the negative-sense strand of to-be-detected DNA respectively starting from one end of the to-be-detected DNA. One of every two adjacent single-chain probe bodies is the single-chain probe body designed according to the nucleotide sequence of the positive-sense strand, and the other one is the single-chain probe body designed according to thenucleotide sequence of the negative-sense strand. The to-be-detected DNA for every two adjacent single-chain probe bodies is not overlapped. The single-chain probe body mixture can cover the whole to-be-detected DNA. The capture probe can remarkably improve the storage capacity of a capture library, the capture efficiency and the effective depth of sequencing data and reduce the repeatability of the sequencing data and quite high in application value.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a capture probe used for high-throughput sequencing to detect gene mutations and an application thereof. Background technique [0002] Whole-genome sequencing can obtain mutations, insertions, deletions, and structural variations at the genome-wide level. However, due to the large capacity of the genome, sequencing at 30× will generate a data volume close to 100G. The low mutation frequency sequencing related to tumors requires at least 1000× coverage, and if the whole genome is sequenced, it will generate as much as 3000G of data. Such a large amount of data will not only cause great difficulties in data analysis, but also generate huge sequencing costs. [0003] The target region liquid phase hybridization capture technology is based on the principle of DNA sequence complementarity, designing specific probes complementary to the target sequence, and synthesizing them o...

AI Technical Summary

Problems solved by technology

The design principles of RNA probes and DNA single-strand probes are basically based on the shingled design method, which is consistent with or opposite to the sense strand of the reference genome, and can only capture libraries consistent with the sense strand or antisense strand of the reference genome part, the other strand is discarded. In this case, if the capture effect is not good, both strands of the library will be lost, reducing the richness of the library.

TWISTbioscience and Mikino use double-stranded probes for library capture, which can ensure that both strands of the library are captured, but because double-stranded probes are reverse complementary structures, many probes will hybridize with each other, resulting in the need to The amount of probe used is large or the capture efficiency is low

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