16S-AmpSeq of mouse stool: MR1 KO Female at 6 months of age
The fecal material was diluted in Cary/Blair media. The Zymo Research Fecal DNA isolation kit was used for all DNA isolations. The isolated DNA was quantitated prior to PCR and then barcoded PCR amplification of the V4 region of the 16S rRNA gene amplicon was accomplished with degenerate primers for use on an Illumina MiSeq sequencer. PCR products were resolved on agarose gels, then DNA was isolated and purified using Qiagen kits, and then quantitated. The products were sequenced on the MiSeq platform, a single flow cell, single lane instrument that generates approximately 9 Gb of sequence data from the paired end 250 bp run.