Amplicon sequencing 16S rRNA V13 of human sample: saliva: Sample CBC.V13.SA.2662754
"Total genomic DNA was extracted from 200mg of fresh stool sample using QIAamp (Qiagen, CA) DNA Stool Mini Kit following manufacturer's instructions. For the construction of pyrotag libraries the V1-V3 hyper variable regions of the 16s rRNA gene was amplified using the 27F (5'AGAGTTTGATCCTGGCTCAG3') and 534R (5'ATTACCGCGGCTGCTGG3') tagged primers. Samples were amplified using two rounds of PCR, the first to amplify the 16S rRNA gene (30 cycles) and a second short round to add barcodes for sample identification (10 cycles), following the procedures detailed in Rascovan et al. 2013. Duplicated reactions were performed in both rounds of PCR to reduce amplification biases. All amplicons were cleaned using Ampure DNA capture beads (Agencourt- Beckman Coulter, Inc.)."