A linker element and method for constructing sequencing library using same

Abstract

Provided is a linker element and a method of using the linker element to construct a sequencing library, wherein the linker element consists of a linker A and a linker B, the linker A is obtained through the complementary pairing of a long nucleic acid strand and a short nucleic acid strand, the 5' end of the long strand has a phosphoric acid modification, and the 3' end of the short strand has an enclosed modification, with enzyme sites in the short strand; the linker B is a nucleic acid single strand, the 3' end thereof can be in a complementary pairing with the 5' end of the long strand of the linker A; and there are II type restriction endonuclease recognition sites in the long strand of the linker A and the linker B. Using the linker element of the present invention for constructing a sequencing library ensures the linking directionality of the linkers while solving the problems of fragment interlinking, linker self-linking and low linking efficiency, and reducing the purification reaction between steps, shortening the linking time and reducing costs.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to an adapter element, a method for constructing a sequencing library using the adapter element, the constructed sequencing library and applications thereof. Background technique [0002] High-throughput sequencing has become one of the foundations of modern molecular biology, biotechnology, medicine and many other fields. In recent years, research on rapid, accurate, and economical methods for determining gene expression levels and nucleotide sequences has continued to innovate; the second-generation high-throughput sequencing technology based on the principle of sequencing while synthesizing has matured. Major sequencing companies have focused on the development of new sequencing products, shortening the sequencing process and reducing costs. The existing sequencing products based on the second generation sequencing technology include whole genome resequencing, whole transcri...

AI Technical Summary

Problems solved by technology

The method for constructing a sequencing library of the present invention abandons the traditional multi-step method of adding adapters at both ends when connecting adapters, and connects adapters with a unique sequence structure and the same novel adapter connection + single-strand replacement. method, while ensuring the directionality of adapter ligation, it solves the problems of fragment interconnection, adapter self-ligation, and low efficiency of fragment ligation, and successfully simplifies the entire adapter ligation process into four new steps, and reduces the purification between steps The reaction greatly shortens the time required for adapter ligation, and significantly reduces the cost; in addition, the sequencing library construction method of the present invention can further introduce nucleic acid probe capture technology, thereby realizing the sequencing of the target genome region, and successfully creating a single-strand library-based Target Region Capture Sequencing Products for Circular Library Sequencing Platforms

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