Resolving Species Level Changes in a Representative Soil Bacterial Community Using Microfluidic Quantitative PCR.

ABSTRACT: Rapid advances in genome sequencing technologies enable determination of relative bacterial abundances and community composition, yet, changes at the species level remain difficult to detect despite importance for certain ecological inferences. We present a method for extraction and direct quantification of species composition of a predefined multispecies bacterial community using microfluidic-based quantitative real-time PCR (qPCR). We employ a nested PCR approach based on universal 16S rRNA gene pre-amplification followed by detection and quantification of absolute abundance of bacterial species using microfluidic array of parallel singleplex qPCR reactions. Present microfluidic qPCR supports 2,304 simultaneous reactions on a single chip, while automatic distribution of samples and react

SUBMITTER: Kleyer H 

PROVIDER: S-EPMC5661172 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature