Absolute quantitative method for microflora based on multi-internal standard system

A microbial community and absolute quantification technology, applied in the field of biotechnology and omics analysis, can solve the problems of inability to quantify biomass, errors, and inability to specifically reflect the law of microbial succession, and achieve shortened sample processing time, reduced errors, and reduced costs. reduced effect
CN110305979AActive Publication Date: 2019-10-08JIANGNAN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
JIANGNAN UNIV
Publication Date
2019-10-08

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Abstract

The invention discloses an absolute quantitative method for microflora based on a multi-internal standard system, which belongs to the field of biotechnology and omics analysis. The present inventionprovides a method for efficiently quantifying microbes during amplicon sequencing, the method is characterized in that the target plasmid with a corresponding gradient is added according to the copy number of the total amount of microbes in a sample, the steps of DNA separation, quality control, and sequencing are carried out to obtain an OTU classification unit, and the absolute number of corresponding microbes in the sample is calculated based on a proportional relationship between the amount of target substance added and the detection amount. The method can realize the absolute quantification of the sample to-be-tested, greatly improves the accuracy of the measurement result, and has the advantages of low cost and the like, and plays an important role in promoting the progress of a sequencing technology.
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Description

technical field

[0001] The invention relates to a method for the absolute quantification of microbial communities based on a multi-internal standard system, which belongs to the field of biotechnology and omics analysis. Background technique

[0002] Amplicon (amplicon) is a nucleotide sequence after DNA or RNA amplification, which is a highly targeted method for analyzing gene variation in specific genomic regions. For example, the ultra-deep sequencing of the amplified fragment of a certain gene obtained by PCR amplification can effectively identify the variation and analyze its characteristics. Amplicon sequencing mainly includes 16S rDNA sequencing, 18S rDNA sequencing, ITS sequencing and target region amplicon sequencing. To reflect the differences between species in the classification of bacteria, fungi, and archaea in environmental samples, it has an important guiding role in the study of microbial composition in environments such as oceans, soils, and intestinal fec...

Claims

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