Sequencing method for increasing reading length

Abstract

The invention relates to the field of gene engineering, and provides a sequencing method for increasing reading length. The method comprises the following steps: reading less than M nucleotide sequences after the anchor extension tail end with enzyme digestion identification site by using a connection sequencing method, and identifying the enzyme digestion identification site carried by the anchor and carrying out enzyme digestion by ingeniously using the cutting endoenzyme; reserving less than N nucleotides after the anchor extension tail end; sealing the positions of the corresponding quantities of nucleotides which have been read; connecting a fluorescence probe for detection after the reserved nucleotides, and reading the information of the nucleotide sequences in the subsequent positions, thereby achieving the goal of increasing the reading length. By using the quantity of the nucleotides reserved after the controlled digestion of the cutting endoenzyme, the invention can ensure the high fidelity of the ligase, thereby ensuring the reading accuracy of the subsequent nucleotide sequence information.

Description

technical field [0001] The present invention relates to the field of genetic engineering, more specifically, to a sequencing method for increasing read length. Background technique [0002] Next-generation sequencing (NGS) mainly includes pyrosequencing (sequencing by pyrosequencing), sequencing by synthesis (SBS) and sequencing by ligation (SBL). Among them, ligation sequencing has the advantages of high accuracy and low cost, and is suitable for transcript and comparative genomics research, especially the detection of single nucleotide polymorphisms (Single Nucleotide Polymorphisms, SNP). [0003] An existing ligation sequencing method is based on oligonucleotide probes with fluorescent labels at specific positions, and the oligonucleotide probes have n bases and are divided into h (h≤n) groups , different fluorescent labels of the same group of oligonucleotide probes correspond to different base sequences at the same specific position, the difference between different gr...

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to provide a sequencing method for increasing the read length, which aims to solve the problem of too short read length of the ligation sequencing method in the prior art, and can further avoid the reading accuracy of nucleotide sequence information when increasing the read length low problem

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