Preparation method and application of nucleic acid library for low host background interference

Abstract

The invention relates to the technical field of amplification of target genes of animals, plants and microorganisms, in particular to a preparation method and application of a nucleic acid library for low host background interference. According to the invention, nucleic acid targeted sequencing analysis technology can be improved, and accurate targeted sequencing, multiple sequencing and high-throughput sequencing can be provided; the nucleic acid library can be used for sequencing low-copy target genes of nucleic acid, and reducing high-abundance nucleic acid interference of a host; the nucleic acid library can be used for targeted sequencing of nucleic acid target genes; and the nucleic acid library can be used for removing nucleic acid target genes for sequencing, and improves detection accuracy.

Description

technical field [0001] The invention relates to the technical field of amplification of target genes of animals, plants and microorganisms, in particular to a nucleic acid library preparation method and application for low host background interference. Background technique [0002] Among the technologies currently involved in biological research, from Sanger sequencing to the second-generation sequencing technology of the Illumina sequencing platform and the third-generation sequencing based on single-molecule sequencing, nucleic acid sequencing technology has been widely used. With the continuous development of sequencing technology, although the sequencing accuracy and precision of different sequencing platforms continue to improve, it is difficult to eliminate the pollution caused by a large number of homologous or non-homologous nucleic acid sequences mixed with the target gene in complex samples This problem cannot be eliminated only from the replacement of sequencing p...

AI Technical Summary

Problems solved by technology

With the continuous development of sequencing technology, although the sequencing accuracy and precision of different sequencing platforms continue to improve, it is difficult to eliminate the pollution caused by a large number of homologous or non-homologous nucleic acid sequences mixed with the target gene in complex samples This problem cannot be eliminated only by changing the sequencing platform

This problem can still be overcome by algorithm optimization and other methods in the sequencing of a single species, but it will bring unavoidable pollution to the mixed sequencing of multiple species (such as metagenomic sequencing)

[0003] In the current methods for sequencing target genes in complex samples, a variety of experimental practices have been improved by using strategies such as nucleic acid purification, polymerase chain reaction, and subsequent removal of sequencing data, but it is inevitable that there will be some changes in the nucleic acid purification kit. Disadvantages such as deviation of sample nucleic acid extraction during use, use of special primers in polymerase chain reaction, partial nucleic acid fragments cannot be amplified, or amplification results deviate from actual abundance, and target genes are covered by high-abundance homologous sequences, etc.

These shortcomings eventually lead to the inability to completely remove nucleic acid sequences with high homology to the target sequence, and the sequence information and abundance of the target sequence cannot be fully obtained, which reduces the accuracy of detection.

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