Dairy white wastewater A
Amplification of the 16S V6-V8  region (16S rDNA) was performed using a two-step dual-indexed PCR approach  specifically designed for Illumina instruments. In a first step, the gene  specific sequence is fused to the Illumina TruSeq sequencing primers and PCR  was carried out. The second PCR step, performed with 50- to 100-fold dilution  of the previously purified PCR product as a DNA template, allowed the  addition of barcodes (dual-indexed) and the missing sequence required for  Illumina sequencing. Barcoded amplicons were then pooled in equimolar  concentration for sequencing on the Illumina Miseq.