Effect of Fecal Microbiota Transplantation on Gut Bacterial Bile Acid Metabolism in Humans
Fecal samples were shipped frozen to the George Washington University Ge-nomics Core for processing. Each sample underwent DNA and RNA extraction in par-allel from 250 mg of fecal material using a ZymoBIOMICS DNA/RNA Miniprep Kit. Resulting DNA was quality controlled using a Thermo Fisher Qubit 3.0 High Sensitivity DNA kit and standardized to 0.2 ng/uL for library construction. Sequencing libraries were prepared, along with a ZymoBIOMICS Microbial Community DNA standard, using a Nextera XT DNA Library Prep kit following Illuminas recommended guide-lines. Libraries were quality controlled using a Thermo Fisher Qubit 3.0 High Sensitivity DNA kit and an Agilent 2100 Bioanalyzer High Sensitivity DNA kit and were subse-quently sequenced as paired-end, 2x150 bp, using a NextSeq 500 Mid-Output kit, with a 1% phi X sequencing control spike-in.