Bacterial DNA was obtained using the DNeasy Blood &Tissue Kit (QIAGEN). PCR for 16S rRNA gene amplification was performed by using the bacterial-specific primers, 27F (5′-AAGGAGGGGATCCAGCCGCA-3′) and 1492R (5′- GTGCCAGCAGCCGCGG -3′). PCR amplifications were performed with KOD plus polymerase as described before39 (link). The PCR product was purified using Wizard PCR Preps DNA Purification System (Promega, Madison, WI, USA). Purified double-stranded PCR fragments were directly sequenced with Big Dye Terminator Cycle sequencing kits (Applied Biosystems, Forster City, CA, USA).
Free full text: Click here
