title: GSM3502727: T3 Replicate 1; Nitrospira moscoviensis; RNA-Seq
library_construction_protocol: Transcriptome analyses were performed using three technical replicates for each of the three sampling points. Biomass of 70 ml culture volume was harvested by centrifugation at 20,000 g, 4 °C, 10-20 min. RNA was immediately extracted using TRIzol reagent (Thermo Fisher Scientific) following the manufacturer's protocol with the following modifications: all incubation and centrifugation times were doubled, RNA was precipitation overnight at –20 °C and afterwards centrifuge at 12,000 g, 4 °C, 20-30 min to collect RNA, no heating step for RNA resuspension was performed. Residual genomic DNA was removed with a DNase I (Thermo Fisher Scientific) treatment following the manufacturer's protocol. Purity of RNA was confirmed on an ethidium bromide stained 0.9% agarose gel. The MEGAclear kit (Thermo Fisher Scientific) was used following the manufacturer's protocol to purify the RNA and remove small RNAs. RNA quantity was controlled using the Qubit RNA HS Assay Kit (Thermo Fisher Scientific). Next, ribosomal RNA was depleted using the MICROBExpress kit (Thermo Fisher Scientific) following the manufacturer's protocol. Transcriptome libraries were prepared with the Ion Total RNA-Seq Kit v2 (Ion Torrent by Thermo Fisher Scientific). RNA yield and size distribution were analyzed using a 2100 bioanalyzer (Agilent Technologies, Santa Clara, CA, USA).
attributes.tag: GEO Accession
attributes.value: GSM3502727