16S V3-V4 Seq of human stool samples
Libraries targeting the V3-V4 hypervariable region were prepared from approximately 12.5 ng of DNA using primers pair: Bakt_341F (CCTACGGGNGGCWGCAG) and Bakt_805R (GACTACHVGGGTATCTAATCC). To reduce chimera formation 23 PCR cycles were carried out (Eppendorf Mastercycler pro; Eppendorf SE, Germany). The Illumina Nextera XT v2 Index-sets (Illumina Inc., USA) were used to barcode the samples. Samples were  pooled equimolarly to a target molarity of ~4 nM. Pool concentration was assessed with QubitTM (Invitrogen, USA) and was sequenced on the Illumina MiSeq platform using a MiSeq Reagent Kit v3 (2 x 300 bp) with a final loading concentration of 8 pM and a PhiX spike-in of 25%.
SUB16216768
PC024