title: gut metagenome of mus musculus
design: Isolated DNA was quantified using the Picogreen system. Primers to amplify the bacterial 16S rRNA gene region was barcoded to label each sample as described previously [Bushman et al 2008].  PCR reactions were carried out in triplicate using Accuprime (Invitrogen, Carlsbad, CA, USA).  Each reaction contain 50 nanograms of DNA and 10 pM of each primer.  Primers annealing to the V1V2 region of the 16S bacterial gene were used for amplification as described previously [Wu et al 2010]. Amplified 16S rDNA will be purified using a 1:1 volume of Agencourt AmPure XP beads (Beckman-Colter, Brea, CA, USA).  The purified products from the stool samples were pooled in equal amounts and analyzed by using the Illumina MiSeq.