Preparation method for DNA library and kit

Abstract

The invention provides a method for preparing a DNA library and specifically capturing and a kit, and the method is suitable for constructing the DNA library of cfDNA, white cell gDNA and DNA from a tissue sample as well as hybridizing and capturing a special target gene area. In the preparation method for the DNA library, terminal repair and 3' end A addition are performed in the same reaction system, and hybridization input is improved, and a blocking preparation in hybridizing and capturing is optimized, so that higher capturing efficiency, higher library variety and more uniform coverage degree are obtained, and therefore, the preparation method is especially suitable for constructing a trace sample library with low-abundance mutation, and is beneficial for realizing saving in technical process. The method provided by the invention can be used for conveniently realizing all variations such as mutation, deletion, insertion, fusion as well as detection on copy number amplification of a target gene, and overcomes the defects that target gene sequences are enriched through a PCR method on DNA level at present, mutation and deletion of the target gene only can be detected, and fusion cannot be detected.

Description

Technical field [0001] The invention relates to a DNA library preparation and targeted capture method and kit. In particular, it relates to the preparation of plasma cell-free DNA library, leukocyte gDNA library or tissue-derived DNA library, and a targeted capture method and kit. Background technique [0002] Studies have found that there is a small amount of free DNA in the blood of cancer patients, namely plasma free DNA (cfDNA), and some of these free DNA are derived from tumor cells. This part of the tumor genome fragment free in the blood is called circulating tumor DNA. (ctDNA). As a new tumor marker, ctDNA plays an important role in individualized drug detection, recurrence and efficacy monitoring of tumors. The method of obtaining cancer-related gene mutation information by detecting the ctDNA released from the tumor tissue into the blood is called "liquid biopsy". In recent years, the concept of liquid biopsy based on ctDNA has been gradually accepted and recognized ...

AI Technical Summary

Problems solved by technology

The method of the present invention can facilitate the realization of all variant types of the target gene, such as mutation, deletion, insertion, fusion, and detection of copy number amplification, and overcomes the current method of enriching the target gene sequence by PCR at the DNA level, but only It can detect the mutation and deletion of the target gene, but cannot detect the disadvantage of fusion

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