v4-16s rRNA sequencing of mouse gut metagenome: Strain_B6;Litter_B;Day_P35;Age_Pup
DNA was extracted using phenol:chloroform extraction followed by clean-up using the DNeasy Blood and Tissue Kit (Qiagen). Amplicons were generated using barcoded PCR primers targeting the V4 region of the 16S rRNA gene as described previously, with 26 cycles of PCR, and purified using Agencourt Ampure XP beads. Maternal body site samples were processed in the same manner as fecal pellets, except 30 cycles of PCR were run to amplify the V4 region. Maternal body site samples were pooled and sequenced separately from fecal samples to ensure adequate sequencing coverage for these low biomass sites.