16S Amplicon Sequencing (Cloudbreak Freestyle™)

Cloudbreak Freestyle chemistry was used to perform amplicon sequencing of the 16S ribosomal (rRNA) gene. 

Methods 

The V3 and V4 regions of bacterial 16S rRNA genes were PCR-amplified from a ZymoBIOMICS Microbial Community DNA Standard using KAPA HiFi HotStart ReadyMix. IDT xGen indexing primers were added using amplification. Libraries were pooled as a 64-plex and sequenced on the AVITI™ System using Cloudbreak Freestyle chemistry with a 2 x 300 read length. Bases2Fastq was used to generate FASTQ files.