454 GS FLX Titanium sequencing; T1CD21S09
The combined V4-V6 variable regions of the 16S rRNA gene were sequenced using the 454 FLX Titanium chemistry.
DNA was extracted from lyophilised caecal content. The V4-V6 16S PCR products were generated with the 16S-0515F(5â-TGYCAGCMGCCGCGGTA-3â) and 16S-1061R (5â-TCACGRCACGAGCTGACG-3â) primers, tailed with Roche MID barcodes 1-72 each side. PCR products were pooled by trial. Roche rapid library adapters we blunt ligated prior to sequencing with the Titanium chemistry on the GS-FLX. Run analysis was performed with the amplicon protocol.