GSM2942996
GSM2942996: T2_60: Treated  Rep2_60min; Caldicellulosiruptor bescii; RNA-Seq
Briefly, cell pellets from each sample were incubated in 250 µL of 20 mg/mL Lysozyme (Sigma Aldrich part number L-7651, St. Louis, MO) resuspended in SET buffer (50 mM Tris-HCl pH 8.0 50 mM EDTA, 20% w/v Sucrose) followed by incubation at 37oC for eight minutes, vortexing briefly every two minutes.  After incubation, 900 µL of buffer RLT from a Qiagen RNeasy Kit (Qiagen, Hilden, Germany) was added to the lysozyme cell mixture and vortexed.  Then 650 µL of 100% ethanol was added to each aliquot and mixed by pipetting.  Each sample was then applied to a RNeasy spin column (Qiagen, Hilden, Germany) and processed following the manufacturers protocol including the optional on column Dnase I treatment.  Samples were eluted in 35 µL RNAse free water (Qiagen, Hilden, Germany). The quantity and quality of RNA was assessed post-elution using a NanoDrop ND-1000 spectrophotometer (NanoDrop Technologies, Wilmington, DE) and Agilent Bioanalyzer (Agilent, Santa Clara, CA, USA). Ribosomal RNA was depleted from the samples using a RiboZero rRNA Removal Kit for bacteria (Illumina Inc. San Diego, CA) cDNA libraries were prepared and barcoded using a ScriptSeq v2 RNA-Seq library preparation kit (Illumina Inc. San Diego, CA) following the manufacturers protocols.
GEO
GSM2942996
GEO Accession
GSM2942996