16S sequencing of cecum from mus musculus
Profiling of the microbiome was completed as described previously using mouse cecum (Org et al., 2015). DNA was isolated from approximately 75 mg of cecum using the DNeasy PowerSoil Kit (Qiagen, Germantown, MD, USA) following the manufacturers instructions. Isolated DNA was checked for concentration using a Nanodrop spectrophotometer. 16S rRNA V4 region primers were used during PCR amplification (Caporaso et al., 2012). The primer sequences used without linker, pad, barcode, or adapter are Forward: GTGCCAGCMGCCGCGGTAA and Reverse: GGACTACHVGGGTWTCTAAT. PCR products were verified, quantified, and pooled before a final clean up using UltraClean PCR Clean-Up Kit before subsequent sequencing on an Illumina HiSeq 3000.