The impact of storage buffer, DNA extraction method, and polymerase on microbial analysis.

ABSTRACT: Next-generation sequencing approaches used to characterize microbial communities are subject to technical caveats that can lead to major distortion of acquired data. Determining the optimal sample handling protocol is essential to minimize the bias for different sample types. Using a mock community composed of 22 bacterial strains of even concentration, we studied a combination of handling conditions to determine the optimal conditions for swab material. Examining a combination of effects simulates the reality of handling environmental samples and may thus provide a better foundation for the standardization of protocols. We found that the choice of storage buffer and extraction kit affects the detected bacterial composition, while different 16S rRNA amplification methods only had a minor e

SUBMITTER: Hallmaier-Wacker LK 

PROVIDER: S-EPMC5908915 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature