Fig. 1

Schematic representation of 16 S amplicon sequencing workflow. E. coli/mock DNA was amplified for 16 S V3-V4 regions using pool of ‘N’ spacer-linked primers with overhang illumina adapters. The amplicons were subjected to second PCR using Nextera XT V2 index primers. Illumina MiSeq platform was used for sequencing the final libraries. The first 100 sequencing cycles illustrates the base diversity generated within a sample amplified using ‘N’ spacer-linked primers