Quantitative 16S metagenome sequencing method

Abstract

The invention discloses a quantitative 16S metagenome sequencing method. Through the adoption of the method of combining random labels with an internal reference method, the accuracy of microbial community structure detection can be effectively enhanced, influences of experiment operation on results can be reduced, and the comparability between sequencing and other molecular biological methods canbe increased; and therefore, the method has important research values for microbial diversity research and unknown pathogen detection.

Description

technical field [0001] The invention relates to the field of sequencing, in particular to a quantitative 16S metagenomic sequencing method. Background technique [0002] 16S rDNA is the most useful and commonly used molecular clock in the taxonomic study of bacteria. It has few types and high content, and exists in all organisms. The 16S rDNA gene sequence includes 9 variable regions and 10 conserved regions. The sequence of the conserved region reflects the relationship between species, while the sequence of the variable region can reflect the difference between species. By designing primers in the two conserved regions of 16S rDNA, the corresponding regions of all bacteria in the sample can be amplified, and because the size of the amplified fragment is moderate, its sequence can be obtained relatively easily by sequencing technology, and the sequencing results can reflect different bacterial genera The difference between them can be compared with 16S databases (such as R...

AI Technical Summary

Problems solved by technology

Since the methods adopted by different research topics are not completely consistent, and the deviations introduced by experimental factors and the actual situation are also different, the comparability of the 16S metagenomic sequencing results between different projects is poor. The data of the 16S metagenomic sequencing project published in China cannot be directly applied to its own research topics and testing projects

At the same time, since the 16S metagenomic sequencing data currently only reflect the relative content (ie percentage), the results cannot be directly compared with the results of traditional culture methods, quantitative PCR methods, and digital PCR methods.

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