RNA-seq of sludge metagenome: Birmingham floc 01
DNA was extracted from sewage sludge using the DNeasy PowerSoil Pro Kit (Qiagen, Manchester, United Kingdom) and purified by using NucleoSpinTM gDNA Clean-up kit (Macherey-Nagel GmbH & Co. KG, Dren, Germany. The primer pro341F (5-CCTACGGGNBGCASCAG-3) and pro805R (5-GACTACNVGGGTATCTAATCC- 3) was used in a target specific PCR target V3-V4 region of the gene encoding the 16S rRNA subunit. The second PCR was followed, in which index sequences were tagged into each amplicon sequence to allow demultiplexing. The PCR products were normalized, pooled, and sequenced on a 2x300 bp paired-end sequencing run using the Illumina MiSeq.