title: 16S rRNA metabarcoding of microbial communities associated with T. reniformis
design: Amplification of the V3-4 region of the 16S rRNA gene was conducted using the illumina-pro341F and illumina-pro805R primer pair. Amplicon PCR reactions were carried out using NEBNext Ultra II Q5 Master Mix with 5ng/l DNA, 1M illumina-pro341F forward primer and 1M pro805R reverse primer followed by PCR clean-up using AMPure XP beads to purify amplicon sequences from primers and primer dimer species. Illumina sequencing adapters and dual indices were attached using the Nextera XT Index Kit followed by a second PCR clean-up using AMPure XP beads. The final DNA library was quantified, normalized and pooled. Pooled libraries were denatured with NaOH, diluted with hybridization buffer and heat denatured before MiSeq sequencing with reagent kit v3 and a 5% PhiX as spike-in.