Amplicon sequencing of prokaryotes: bacterial and archaeal 16S rRNA (V3V4) from endosymbiont samples - Rep3
Microbial community composition was assessed by targeted amplification of the 16S rRNA gene from environmental DNA samples. The V3V4 hypervariable regions of bacterial and archaeal 16S rRNA genes were amplified using domain-specific primers in a first PCR with variable input DNA and cycle number. A second PCR was performed to incorporate Illumina sequencing adapters and sample-specific barcodes. Amplicons were quality-checked, quantified, pooled in equimolar concentrations, and prepared for paired-end sequencing.