Sequencing library building method and reagent based on molecular inverse probe

Abstract

The invention discloses a sequencing library building method and reagent based on a molecular inverse probe. The method includes: using the molecular inverse probe and denatured nucleic acid to perform annealing hybridization, wherein the molecular inverse probe comprises the anchoring sequence of a 5' end, the extension sequence of a 3' end and the sequencing connector sequence between the anchoring sequence and the extension sequence, and the anchoring sequence and the extension sequence are respectively inversely complementary with sequences at two ends of a target area; using the target area as a template, and performing polymerization reaction starting from the extension sequence of the 3' end to generate the supplementary sequence of the target area; connecting the anchoring sequence of the 5' end with the supplementary sequence of the target area to form cyclic annular nucleic acid molecules; using exonuclease to digest non-cyclized linear molecules to obtain single-chain cyclic annular molecules containing the target area; sequencing the obtained single-chain cyclic annular molecules so as to achieve capture sequencing of the target area. The sequencing library building method has the advantages that the building of a single-chain cyclic annular library and target area capture are integrated, and library building process and cycle are shortened greatly.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a molecular reverse probe-based sequencing library construction method and reagents. Background technique [0002] With the development of DNA sequencing technology, high-throughput sequencing technology has been widely used in various fields of life science research. Although the cost of sequencing technology is getting lower and lower with the continuous updating and popularization of sequencing technology, the cost of whole genome sequencing technology itself is still very expensive. A better solution to reconcile this problem is to enrich the target region of interest for high-throughput sequencing. Traditional sequence capture technology usually constructs a high-throughput sequencing library, and then uses probes to enrich the library of the target region and then sequence. [0003] The sequencing platform represented by the CG (Complete Genomics) sequencing pla...

AI Technical Summary

Problems solved by technology

[0005] At present, there is no application of MIP technology in the sequencing platform with single-stranded circular library as the sequencing object, to solve the problem of time-consuming, labor-intensive, cost-consuming and complex enrichment of target regions in the construction of single-stranded circular library

Images