S4 Bacteria
Library preparation was performed at Genotypic Technology’s Genomics facility. 4ng of Nanodrop quantified DNA was used for amplifying V3-V4 region of 16S region with specific primers which also have a ‘tag’ sequence that are complementary to Illumina sequence adapter and index primers from the Nextera XT Index kit V2. This round of PCR generates single Amplicon of ~480-530 bp. The Amplified products were checked on the Agarose Gel before proceeding for Indexing PCR. In the next round of PCR (indexing PCR) Illumina sequencing adapters and dual Indexing barcodes are added to 25ng of round1 product using limited cycle PCR to give a final product of ~580-600 bp. The library was diluted (1:10) and validated for quality by running an aliquot onHigh Sensitivity Tape Station(Agilent).