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Method for performing sequencing and cluster analysis on V6 hypervariable region of metagenomic 16S rDNA

A technique of metagenomic and clustering analysis, which is applied in the field of sequencing and clustering analysis of metagenomic 16S hypervariable region V6, can solve the problems of insufficient sequencing information, microbial classification, short read length, and inability to sequence 16S rDNA, and saves economic costs. , the effect of reducing human labor

Inactive Publication Date: 2012-05-30
BGI SHENZHEN CO LTD +1
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Problems solved by technology

A major challenge for these revolutionary technologies is that the read lengths are too short to sequence the 16S rDNA of each individual, and thus its sequencing information is not sufficient to allow us to accurately classify microorganisms

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  • Method for performing sequencing and cluster analysis on V6 hypervariable region of metagenomic 16S rDNA
  • Method for performing sequencing and cluster analysis on V6 hypervariable region of metagenomic 16S rDNA
  • Method for performing sequencing and cluster analysis on V6 hypervariable region of metagenomic 16S rDNA

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Embodiment Construction

[0022] The present invention will be described more fully hereinafter with reference to the accompanying drawings, in which exemplary embodiments of the invention are illustrated.

[0023] figure 1 A flow chart showing a method for performing sequencing cluster analysis on the 16S hypervariable region V6 of the metagenomics provided by an embodiment of the present invention.

[0024] Such as figure 1 As shown, the method flow 100 for sequencing and clustering analysis of the metagenomic 16S hypervariable region V6 includes:

[0025] Step 102, extracting the deoxyribonucleic acid DNA of the microorganism. For example, the Ultraclean Soil DNA kit (MoBio, USA) is used to extract microbial DNA from the sample sediment.

[0026] Step 104, the hypervariable region V6 of the metagenomic 16S ribosomal deoxyribonucleic acid rDNA by primers (the two ends of this region each have a conserved region of about 20 base pairs bp, and the variable region in the middle is about 60-90bp) Per...

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Abstract

The invention discloses a method for performing sequencing and cluster analysis on V6 hypervariable regions of metagenomic 16S rDNAs. The method comprises the following steps of: extracting DNAs of microbes; performing polymerase chain reaction (PCR) on V6 hypervariable regions of metagenomic 16S rDNAs, and inserting a tag sequence in each sample; hybridizing PCR products of different samples; setting a database of hybrid PCR products by Solexa method; performing pair-end sequencing on the library of V6 hypervariable regions by using Solexa sequencing tools to obtain primary sequencing data; screening the sequencing data to filter low-quality data; assembling full-length sequences of the V6 hypervariable regions by means of contig relationship; distributing reads to the corresponding samples by the tag sequences; and performing classification analysis on the reads to complete high-throughput and accurate classification of microbial populations by sequencing the hypervariable regions.

Description

technical field [0001] The invention relates to the technical field of microbial gene sequencing and analysis, in particular to a method for sequencing and clustering analysis of the metagenome 16S hypervariable region V6. Background technique [0002] In order to study the types of microbial populations in the biological environment, the general traditional methods include: direct cultivation of microorganisms, denaturing gradient gel electrophoresis (DGGE, Denaturing Gradient Gel Electrophoresis), terminal restriction endonuclease fragment length polymorphism (T- RFLP, Terminal Restriction Fragment Length Polymorphism), fluorescence in situ hybridization (FISH, Fluorescence In Situ Hybridization), PCR (polymerase chain reaction, Polymerase Chain Reaction) on possible microbial species; but these methods can only reveal the A very small number of microbial species. If metagenome analysis can be carried out, a relatively comprehensive catalog of microbial species can be obt...

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/689
Inventor 刘晓周宏伟栗东芳
Owner BGI SHENZHEN CO LTD
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