DNA/RNA seq of mixed culture: biocathode of EMC
DNA/cDNA samples by PCR amplification with universal primers for prokaryotic 16S rRNA gene sequences, 515F and 806R, were used to construct libraries for next-generation sequencing (NGS). An Illumina adapter sequences were added to the 5  end of each primer. PCR was conducted with the following thermal cycling profile: an initial denaturation at 94oC for 10 min, 30 cycles of amplification (30 s at 94oC, 30 s at 55oC, and 30 s at 72oC), and a final extension at 72  C for 7 min. The PCR products were submitted to Macrogen, Inc. (Korea) for NGS on the Illumina MiSeq platform. Reads with low quality scores (<20), ambiguous bases, or potential chimeric sequences were excluded. The trimmed sequences were aligned and clustered using CD-HIT-OTU (http://weizhongli-lab.org/cd-hit-otu/) with an operational taxonomic unit (OTU) definition of  97% sequence similarity (i.e., <3% divergence). Taxonomic classification was performed using the QIIME pipeline against the SILVA database.
SUB3399125
ABLE