Construction method and application of high-throughput sequencing library

Abstract

The invention discloses a construction method and application of a high-throughput sequencing library. The method for constructing the high-throughput sequencing library comprises the steps that genomic DNA is fragmented, a basic group A is added to the 3'end of a DNA fragment subjected to end repairing, the DNA fragment is connected with joints, amplification is conducted, then a DNA library is mixed with annular closed oligonucleotides and a specific probe so as to conduct hybrid capturing, and the introduced joints and an introduced tag sequence are closed by the annular closed oligonucleotides, wherein the annular closed oligonucleotides are designed for the joints and the tag sequence, and the annular closed oligonucleotides and the joint sequences at the two ends of the library sequence are subjected to complementary pairing; and a connecting product is subjected to hybrid capturing through the specific probe so as to obtain a target fragment. According to the construction methodand application of the high-throughput sequencing library, a DNA template closing the sequencing joint sequences in an annular mode is captured, and the efficiency of capturing the target DNA sequence by the probe is improved.

Description

technical field [0001] The present invention relates to the field of biotechnology. In particular, it relates to targeted sequencing techniques involving the determination of target DNA fragments of a sample. More specifically, the present invention provides a method for constructing a high-throughput sequencing library, a sequencing method for determining a target DNA fragment of a sample, a device for determining a sample target DNA fragment, and a method for constructing a sample target DNA fragment. A kit for high-throughput sequencing libraries of DNA fragments. Background technique [0002] The new-generation high-throughput sequencing technology that has emerged in recent years can simultaneously sequence billions of DNA fragments, providing a powerful tool for basic biomedical research and clinical testing. Whole-genome sequencing is widely used in the field of basic scientific research due to its comprehensive and comprehensive detection performance. However, the ...

AI Technical Summary

Problems solved by technology

[0005] However, the target-targeted sequencing technology based on probe capture has a long library construction process, and it takes 1-2 days or even longer for the probe to fully bind to the target region, which greatly limits the timeliness of clinical testing.

In addition, the efficiency of hybrid capture is limited (usually only 50-60% capture efficiency), and the data wasted in non-target regions also virtually increases the cost of probe capture

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