Method and kit for rapidly constructing target nucleic acidsequencing library on basis of probe capture and enrichment

Abstract

The invention discloses a method and a kit for rapidly constructing a target nucleic acidsequencing library on the basis of probe capture and enrichment. The method comprises following steps and adopts reagents required for the steps: genome DNA is subjected to fragmentation; DNA end repair is performed; a probe and a single-stranded DNA fragment formed through thermal denaturation are subjected to molecular hybridization; biotin-avidinmagnetic bead separation is executed; the probe is taken as a primer for primer extension, and a complementary strand is synthesized; one end of a double-stranded DNA fragment is connected with an linker, and the other end is not connected with the linker; PCR amplification is performed; DNA fragment lengthselection is executed; the reagents are used for completing the reactions of all the steps. Compared withconventionaltarget nucleic acid enrichment technologies, the method has the advantages of good fidelity, good reproducibility, high speed and high universality.

Description

technical field [0001] The invention belongs to the technical field of high-throughput gene sequencing. technical background [0002] Compared with classic Sanger sequencing, high-throughput gene sequencing is also called next-generation sequencing or next-generation sequencing (NGS). Correspondingly, Sanger sequencing is also called generation sequencing. Whole exome sequencing and panel sequencing are both targeted gene sequencing, which is an important branch of next-generation sequencing technology. %, the target area targeted by various panels is smaller; the second is to reduce the difficulty of bioinformatics data analysis and speed up the analysis, so it is widely used in various fields such as basic research, clinical research, clinical diagnosis and new drug development. [0003] There are currently four main brands of exome sequencing and targeted sequencing products: SureSelect and HaloPlex from Agilent, NimbleGen from Roche, TruSeq and Nextera from Illumina (b...

AI Technical Summary

Problems solved by technology

Among them, although the Nextera technology based on transposons is fast, the data quality is slightly flawed

Other exome sequencing technologies include the construction of the whole genome library and the capture and enrichment of exon region fragments. The steps are numerous and time-consuming; due to the many steps of reaction and purification, the loss of DNA is relatively large, so the template The amount of genomic DNA is relatively high

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