Metagenome Sequencing and Analysis of 16s rRNA V3-V4 region: SO_9612_20
Genomic  DNA  was  amplified  for  26  cycles  using  KAPA  HiFi  HotStart  PCR  Kit.  The  forward  and  reverse  primer  concentration  was  5uM each. The amplicons were analyzed on 1.2% agarose gel. 1l  of  diluted  round  1  PCR  amplicons  were  used  for  Indexing  PCR.  Here,  the round 1 PCR amplicons were amplified for 10 cycles to add  Illumina sequencing barcoded adaptors (Nextera XT v2 Index Kit). Round 2 PCR amplicons were  analyzed  on  1.2%  agarose  gel.  Further  the  libraries  were  normalized  and pooled for multiplex sequencing.