The effect of reverse transcription enzymes and conditions on high throughput amplicon sequencing of the 16S rRNA.

ABSTRACT: It is assumed that the sequencing of ribosomes better reflects the active microbial community than the sequencing of the ribosomal RNA encoding genes. Yet, many studies exploring microbial communities in various environments, ranging from the human gut to deep oceans, questioned the validity of this paradigm due to the discrepancies between the DNA and RNA based communities. Here, we focus on an often neglected key step in the analysis, the reverse transcription (RT) reaction. Previous studies showed that RT may introduce biases when expressed genes and ribosmal rRNA are quantified, yet its effect on microbial diversity and community composition was never tested. High throughput sequencing of ribosomal RNA is a valuable tool to understand microbial communities as it better describes the ac

SUBMITTER: Stovicek A 

PROVIDER: S-EPMC6816399 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature