Fermenter metagenome: F25
Amplification of the hypervariable V4 region of the 16S rRNA gene. Dual-indexed Illumina compatible libraries in a single PCR step. Fusion primers with target-specific portions were used in the first PCR, and Fluidigm CS oligos at their 5' ends. Second, PCR targeting the CS oligos was used to add sequences necessary for Illumina sequencing and unique indexes. PCR products were normalized using Invitrogen SequalPrep DNA normalization plates and the pooled samples were sequenced using an Illumina MiSeq flow cell (v2) in a 500-cycle reagent kit (2x250bp paired-end reads). Finally, sequencing was done.