Gradient Internal Standard Method for Absolute Quantification of Microbial Amplicon Sequencing Data.

ABSTRACT: High-throughput amplicon sequencing is a critical tool for studying microbiota; however, it results only in relative abundance data. Thus, changes in absolute abundance of microbiota cannot be determined, which hinders further microbiology research. We have therefore established a gradient internal standard absolute quantification (GIS-AQ) method to overcome this issue, which can simultaneously obtain the absolute abundances of bacteria and fungi. Deviations from the quantitative equations of microbes and internal standards were eliminated through calibration. Compared with traditional quantitative real-time PCR and microscopy quantifications, this method is reliable (R ² _average = 0.998; P < 0.001) and accurate (P _{internals versus microscopy}

SUBMITTER: Wang S 

PROVIDER: S-EPMC7901480 | biostudies-literature | 2021 Jan

REPOSITORIES: biostudies-literature