title: 16S rRNA gene amplicon sequencing (V3-V4 region)
design: DNA isolation was performed from 25 mg tissue samples or 250 ul urines according to manufacturers protocol by the ZymoBIOMICS DNA Miniprep Kit,  after the enzymatic dissolve with ProtK in 56 C, 3 hours in the case of tissue samples. The concentration of genomic DNA was measured using a Qubit2.0 Fluorometer with Qubit dsDNA HS Assay Kit. Bacterial DNA was amplified with tagged primers covering the V3-V4 region of the bacterial 16S rRNA gene. Amplicons were cleaned with AMPure XP beads. Index PCR  attached dual indices and Illumina sequencing adapters using the Nextera XT Index Kit. Final libary before quantification was cleaned up with AMPure XP beads. Barcoded DNA samples were quantified with Quibit dsDNA HS assay Kit with Quibit2.0, and quantified with DNA 1000 Kit with Agilent 2100 Bioanalyzer. Libaries were normalised, pooled, diluted to 13 pM and denaturated with NaOH prior to sequencing on Illumina MiSeq, using MiSeq reagent kit v3 600 cycle
library_name: E-30v_S93_L001