16S rRNA amplicon sequences of mixtures of stool DNA: E01JH0017  Titration 2^-10
Sample prepared using the Illumina 16S library protocol (16S Metagenomic Sequencing Library Preparation (11/27/2013, https://support.illumina.com). The PCR assay targeted the V3-V5 region of the 16S rRNA gene, Bakt_341F and Bakt_806R. Primer sequences include overhang adapter sequences for library preparation (forward primer 5'- TCG TCG GCA GCG TCA GAT GTG TAT AAG AGA CAG CCT ACG GGN GGC WGC AG - 3' and reverse primer 5'- GTC TCG TGG GCT CGG AGA TGT GTA TAA GAG ACA GGA CTA CHV GGG TAT CTA ATC C - 3'). PCR products were indexed using the Illumina Nextera XT index kits. Sample concentration was normalized using SequalPrep Normalization Plate Kit (Catalog n. A10510-01, Invitrogen Corp., Carlsbad, CA), according to the manufacturer's protocol. Due to the low pooled amplicon library DNA concentration, a modified protocol for low concentration libraries was used. Libraries were sequenced using using the MiSeq 2 X 300 bp Reagent Kit v3. Base calls were made using Illumina Real Time Analysis Software version 1.18.54.