Quantifying and Understanding Well-to-Well Contamination in Microbiome Research.

ABSTRACT: Microbial sequences inferred as belonging to one sample may not have originated from that sample. Such contamination may arise from laboratory or reagent sources or from physical exchange between samples. This study seeks to rigorously assess the behavior of this often-neglected between-sample contamination. Using unique bacteria, each assigned a particular well in a plate, we assess the frequency at which sequences from each source appear in other wells. We evaluate the effects of different DNA extraction methods performed in two laboratories using a consistent plate layout, including blanks and low-biomass and high-biomass samples. Well-to-well contamination occurred primarily during DNA extraction and, to a lesser extent, in library preparation, while barcode leakage was negligible. Lab

SUBMITTER: Minich JJ 

PROVIDER: S-EPMC6593221 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature