16S V3V4-Seq of human stool: gut microbiome
DNA was extracted from human stool using chemagic DNA Stool Kit (PerkinElmer chemagen, Baesweiler, Germany) combined with a previous bashind bead step. A 2 step PCR was used to amplify the 16S rDNA V3V4 region and barcoded using Illumina Nextera Indices. Before proceeding to the next step, the pool was denatured and diluted according to the manufacturer's instructions. Sequencing was performed on an Illumina MiSeq system with the MiSeq Reagent Kit v3, utilizing 2  300 cycles. Clustering was carried out at 6 pM with a 15% Phi-X spike-in.
SUB15292549
IPNV-087