title: Metatranscriptome of alkaline anaerobic reactor
design: 17.91 µg of total RNA were extracted from 16ml alkaline sludge. After DNase and Riboblock RNase inhibitor (Fermentas, Thermo Fisher Scientific GmbH, Deutschland) treatment and purification with Qiagen RNeasy MiniElute Cleanup kit (Qiagen Gmbh, Deutschland), 11.23 µg of purified RNA remained. Ribo-zero rRNA removal kit was used to remove ribosomal RNA.  For cDNA library preparation, the “Ion Total RNA-seq kit v2 for whole transcriptome libraries” was used.  20.61 ng mRNA form the sample were used to construct the 400bp cDNA library.Sequencing template preparation was performed using the OneTouch Instrument and the OneTouch ES module. Enriched ISP particles were sequenced with the Ion PGM™ 400 Sequencing Kit (Life Technologies, USA) on a 318™ Chip with 1000 flows following the manufacturer's instructions. Automated analysis was performed with the Torrent Suite™ Software v3.2 using default settings.