BioProject

The primary 16S rRNA sequencing protocol for microbial community analysis using Illumina platforms includes a single-indexing approach that allows pooling of hundreds of samples in each sequencing run. More...

The primary 16S rRNA sequencing protocol for microbial community analysis using Illumina platforms includes a single-indexing approach that allows pooling of hundreds of samples in each sequencing run. The protocol targets the V4 hypervariable region (HVR) of 16S rRNA using 150bp paired-end (PE) sequencing. However, recent improvement in Illumina chemistry has increased the read length up to 600bp using 300bp PE sequencing. To take advantage of the longer read length, a dual-indexing approach has been developed for targeting different HVRs. However, due to simple working protocols, the single-index 150bp PE approach still continues to be attractive to many researchers.

In this study, we tested an extended single-indexing protocol for 300bp PE Illumina sequencing that targets the V3-V4 HVRs of 16S rRNA.

The test was conducted on 11 digesta samples that were collected from a biogas fermenter fed by feedstock manure. The biogas fermenter samples were purposely selected as they have greater microbiota diversity compared to microbiota in human and animal body sites. Less...