Establishment method of gene library of fecal flora based on high-throughput gene sequencing

Abstract

An establishment method of a gene library of fecal flora based on high-throughput gene sequencing employs a ''nested PCR'' method for enrichment amplification on 16S rDNA, so as to reduce the host and food residue genome pollution by the maximum; and V3 and V6 are combined for specific amplification and massive parallel sequencing, so as to obtain the a target gene sequence database of the flora. The library can increase the identification of the bacterial flora from Genus level to Species level. The method can maximally avoid the interference of host cell nucleic acid, and completely and efficiently detect 16SrDNA tag sequence of all bacteria varieties, so as to accurately determine the ecological structure of the intestinal flora.

Description

technical field [0001] The invention belongs to the technical field of molecular biology detection methods for bacteria in the field of biotechnology, and in particular relates to a method for establishing a fecal flora gene library based on high-throughput gene sequencing. Background technique [0002] In recent years, the relationship between gut microbes, diet and health has been increasingly explained, largely due to the rapid development of gut microbiome research technology. The research on gut microbes has generally gone through three stages: culture-dependent methods, culture-independent traditional molecular biology methods, and sequencing-based high-throughput omics methods. [0003] By comparing the characteristics and applications of the research methods in the three stages, people can find that the culture-dependent method can obtain the corresponding strains while identifying the strains, which is convenient for subsequent research, but the culture method itsel...

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Problems solved by technology

[0007] In traditional molecular biology methods, the Sanger method is sometimes used to sequence and identify 16SrDNA of a single s

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