Flx titanium reagents

Faecal bacterial DNA was isolated according to procedures described previously^{(16 )} using the MO BIO PowerSoil™ Kit (MO BIO Laboratories, Carlsbad, CA, USA). Amplification of a 600 bp sequence of the V4–V6 variable region of the 16S rRNA gene was done using barcoded primers as previously described^{(17 )}. PCR amplicons were further purified utilising AMPure XP beads (Beckman-Coulter Inc.). Amplicons were combined in equimolar ratios to create a DNA pool that was used for pyrosequencing. DNA quality of amplicon pools was assessed before pyrosequencing using a 2100 Bioanalyser (Agilent Technologies). Pyrosequencing was performed at the W. M. Keck Center for Biotechnology at the University of Illinois utilising a 454 Genome Sequencer and FLX titanium reagents (Roche Applied Science).

Faecal bacterial DNA was isolated according to procedures described previously^{(20 )} using the MO BIO PowerSoil™ Kit (MO BIO Laboratories). Amplification of a 600 bp sequence of the V4–V6 variable regions of the 16S rRNA gene was done using barcoded primers as previously described^{(21 )}. PCR amplicons were further purified utilising AMPure XP beads (Beckman-Coulter Inc.). Amplicons were combined in equimolar ratios to create a DNA pool that was used for pyrosequencing. DNA quality of amplicon pools was assessed before pyrosequencing using a 2100 Bioanalyzer (Agilent Technologies). Pyrosequencing was performed at the W. M. Keck Center for Biotechnology at the University of Illinois utilising a 454 Genome Sequencer and FLX titanium reagents (Roche Applied Science).