16s rRNA seq of mus musculus: adult female fecal pellet
Bacterial genomic DNA was isolated using the DNeasy PowerSoil Kit (Qiagen). 50mg per sample, bead-beat in buffer RLT using Lysing Matrix E tubes and extracted using AllPrep DNA/RNA/Protein Mini kit (Qiagen). Sequencing libraries were generated according to methods adapted from Caporaso et al. 2011101, amplifying the V4 regions of the 16S rRNA gene by PCR and purified using the Qiaquick PCR purification kit. 250 ng of purified, PCR product from each individually barcoded sample were pooled and sequenced by Laragen, Inc. using the Illumina MiSeq platform 2 x 250bp paired-end sequencing.