DNA fragment targeted enrichment method and application thereof in genome targeted sequencing

A targeted enrichment and fragmentation technology, applied in the field of cas9-capture, can solve the problems of DNA capture method capture bias, etc., and achieve the effect of simple operation, simple operation, and reduced loss

Inactive Publication Date: 2021-07-13
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0005] The main purpose of the present invention is to provide a DNA fragment targeted enrichment method to solve the technical problem of capture bias in the DNA capture method of the prior art

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  • DNA fragment targeted enrichment method and application thereof in genome targeted sequencing
  • DNA fragment targeted enrichment method and application thereof in genome targeted sequencing
  • DNA fragment targeted enrichment method and application thereof in genome targeted sequencing

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Embodiment Construction

[0038] The technical solutions in the embodiments of the present invention will be clearly and completely described below, obviously, the described embodiments are only some of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

[0039] In addition, the technical solutions of the various embodiments of the present invention can be combined with each other, but it must be based on the realization of those skilled in the art. When the combination of technical solutions is contradictory or cannot be realized, it should be considered as a combination of technical solutions. Does not exist, nor is it within the scope of protection required by the present invention.

[0040] Unless otherwise specified, various raw materials, reagents, instruments and equipment u...

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Abstract

The invention discloses a DNA fragment targeted enrichment method which comprises the following steps: analyzing upstream and downstream guide RNA of a target fragment according to DNA sequence information of the target fragment, and synthesizing the guide RNA; obtaining sample DNA; carrying out in-vitro cas9 enzyme digestion reaction on the sample DNA through a cas9 protein and the guide RNA, so as to cut the target fragment from the sample DNA; carrying out agarose gel electrophoresis on the sample DNA cut by the in-vitro cas9 enzyme digestion reaction, and cutting off a gel block corresponding to the size of the target fragment strip; and recovering the DNA in the gel block and carrying out sequencing. The DNA fragment targeted enrichment method is high in applicability and small in introduced bias.

Description

technical field [0001] The invention relates to the technical field of cas9-capture, in particular to a DNA fragment targeted enrichment method and its application in genome targeted sequencing. Background technique [0002] In molecular experiments, a large number of sequencing steps are required. In most cases, scientific research or clinical testing only focuses on some specific genes on the genome. At this time, the sequencing of the whole genome is actually a waste of most of the sequencing data. Therefore, DNA capture technology is very important for high-throughput sequencing. Widespread application of technology is significant. [0003] At present, there are many technical solutions focusing on DNA capture suitable for next-generation sequencing. Due to the short read length of next-generation sequencing, small fragments of DNA can be captured relatively easily by methods such as probe capture and PCR capture. However, because most of the existing capture methods a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806C12Q1/6869C12N15/10
CPCC12Q1/6806C12Q1/6869C12N15/1003
Inventor 李杰群司中洲谭洁琼
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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