DNA extraction for microbiome studies

Dna Extraction For Microbiome Studies

DNA extraction for microbiome studies

Method for preparing DNA from feces in guanidine thiocyanate solution affects 16S rRNA-based profiling of human microbiota diversity

 2017 Jun 28;7(1):4339. doi: 10.1038/s41598-017-04511-0.

Here, we examined the influence of various factors such as pretreatment (e.g., removing GuSCN solution and washing feces with phosphate-buffered saline (PBS) before mechanical lysis), fecal concentration in the GuSCN solution, storage time, and position of fecal subsampling on the 16S rRNA-based analysis of fecal bacteria in GuSCN solution. We found that pretreatment and fecal concentration affected the bacterial composition, and a little change was noted with subsampling position. Based on these results, we propose a basic protocol, including fecal sampling, sample storage, and DNA extraction, for the 16S rRNA-based analysis of bacterial composition in feces suspended in GuSCN solution.

CONSORTIA

Telethon KIds Institute
DOHaD ANZ
Australian Phenomics Facility
Lottery West State Biomedical Facility Genomics
Centre for Comparative Genomics
Australian Genome Research Facility
WA Human Microbiome Collaboration Centre
WAHTN
West Coast Microbiome Network