title: GSM2879505: T60-None-2; Clostridium acetobutylicum ATCC 824; RNA-Seq
library_construction_protocol: Total RNA was isolated using the miRNeasy Mini Kit (Qiagen; 217004) according to the manufacturer's protocol, with an additional homogenization and mechanical disruption step using a bead beater (BioSpec, Bartlesville, OK, USA) with Zirconia/Silica beads (BioSpec; 11079101z). RNA quality was assessed using the 2100 BioAnalyzer (Agilent Technologies, Santa Clara, CA, USA) and quantified using a spectrophotometer (DeNovix, Wilmington, DE, USA) and was stored at -80°C until DNase treatment. DNA was removed using the TURBO DNA-free kit (Life Technologies; AM1907) according to the manufacturer's protocol. RNA was quantified and quality assessed, as stated above, and genomic DNA depletion was confirmed using qPCR with 16S rRNA gene primers [19] and iQ SYBR Green Supermix (Bio-Rad; 170-8882) according to the manufacturer's instructions. Ribosomal RNA was removed using the Ribo-Zero rRNA Removal Kit (Gram-Positive Bacteria) (Illumina; MRZGP126) according to the manufacturer's protocol. The quality of rRNA depleted samples was assessed using a 2100 BioAnalyzer prior to processing for sequencing library generation. TruSeq Stranded mRNA Sample Preparation Kit (Illumina; RS-122-2101) was used to prepare the rRNA depleted RNA for sequencing according to the manufacturer's protocol. Libraries were quantified using the Kapa Library Quantification Kit (KapaBiosystems; KK4854) according to the manufacturer's instructions, then normalized and pooled for sequencing according to the Denature and Dilute Libraries Guide for the NextSeq 500 (Illumina; Doc No – 15048776 v02).
attributes.tag: GEO Accession
attributes.value: GSM2879505