Metagenome of gut microbiota of Wistar rat
DNA was isolated from approximately 200 mg of stool using two different commercially-available kits: QIAamp DNA Stool Minikit (Qiagen) and MoBio PowerSoil DNA Isolation Kit (MoBio Laboratories). A barcoded pyrosequencing approach was employed for in-depth analysis of the bacterial community composition and diversity. Briefly, the V1-V3 and V3-V5 hypervariable regions of bacterial 16S rRNA gene were PCR-amplified with the primer pairs 27F-534R and 357F-926R, respectively, according to the ÒHuman Microbiome Project (HMP) 454 16S Protocol Version 4.2.2. The primer pair systems generated amplicons of around 500 and 560 bp in length for V1-V3 and V3-V5 regions, respectively. The samples were sequenced following the abovementioned protocol. The structure was forward primer-B adapter-barcode-reverse primer-A adapter. Bacterial amplicons were delivered for pyrosequencing on a 454 Genome Sequencer GS FLX Titanium platform (Roche Diagnostics Ltd).