Microbial community composition method and device based on high-throughput sequencing

Abstract

The invention discloses a microbial community composition method and device based on 16S rRAN gene hypervariable regions V1 / V2. The method comprises the steps of extracting DNAs in microbe samples and carrying out PCR (Polymerase Chain Reaction) amplification on the hypervariable regions V1 / V2 of the 16S rRANs of the DNSs in the samples; carrying out Solexa database establishment on amplified products and marking each sample through junctions with label sequences in a database establishment process; mixing different samples with the label sequences, carrying out sequencing on the mixed samples through utilization of a Solexa sequencing tool, thereby obtaining original sequencing sequence reads distinguished according to labels; carrying out assembling through utilization of overlapping relationships of the reads, thereby obtaining unique reads of the hypervariable regions V1 / V2 or V6; and carrying out sequence similarity comparison analysis on the unique reads, thereby classifying microbes in the samples and realizing relatively abundant calculation. According to the method and the device provided by the invention, the microbial community composition is analyzed accurately.

Description

technical field [0001] The invention relates to the technical field of bioinformatics analysis, in particular to a method and a device for classifying the V6 region of the 16S rRNA hypervariable region of the microbial genome. Background technique [0002] Traditional methods for the type and abundance of microbial populations include: direct culture of microorganisms, denaturing gradient gel electrophoresis, terminal restriction endonuclease fragment length polymorphism, fluorescence in situ hybridization, and PCR for possible microbial species (polymerase chain reaction); but these methods can only reveal a small part of the microbial species in the environment. If metagenome analysis can be carried out, a relatively comprehensive catalog of microbial species can be obtained by directly conducting genome research on microbial populations in the environment, which will help the follow-up research and application of microbial populations. [0003] The sequence of 16S rRNA (...

AI Technical Summary

Problems solved by technology

A major challenge for these revolutionary technologies is that the read lengths are too short to sequence each individual 16S rRNA, and thus its sequencing information is insufficient to allow us to accurately classify microorganisms

However, the determination of the variable region of 16S rRNA can be used to classify microorganisms, by designing specific universal primers to perform specific PCR (polymerase chain reaction, Polymerase Chain Reaction) on the 16S variable region, and then sequenced with a sequencer, established in The phylogenetic tree on this method shows good biodiversity, but its sequencing cost is high, although it is 1 / 10 of the cost of traditional capillary sequencing, it is about 10 times the cost of other next-generation sequencers

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