Construction method and application of single-stranded sequencing library

Abstract

The invention discloses a construction method and application of a high-throughput sequencing library, wherein the construction method of the high-throughput sequencing library includes the steps: carrying out fragmentation of genomic DNA and adding a base A to the 3' end of an end-repaired DNA fragment, connecting with a connector and amplifying, and digesting the DNA library with exonucleases toobtain a single-stranded DNA library; and carrying out hybrid capturing of the connected product by a specific probe, so as to obtain a target fragment. The DNA hybridization process is improved, theconventional double-stranded DNA template is digested into a single strand by enzymatic digestion, then the introduced connector and tag sequence are completely sealed by cyclic oligonucleotides, andthe single-stranded DNA template is captured with a probe (RNA or DNA); therefore, the time of hybrid capturing can be shortened, the efficiency of capturing the target DNA sequence by the probe is improved, and the preference of GC regional capturing is reduced.

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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