title: WT_3_4H
organism: Clostridium beijerinckii NCIMB 8052
description: Protocols: Total RNA was isolated from C. beijerinckii NCIMB 8052 for transcriptome studies. Samples were taken from biological triplicate of 400-mL fermentations from cells in early exponential, acetogenic, and solventogenic phase. Cells were pelleted for 15 min at 3000 × g and 4 °C, washed 3 times with sterile H2O and stored at -80 °C until further use. Fermentations were performed at 35 °C in 400 mL working volume of CM2 medium containing (in g L-1): yeast extract, 1.00; KH2PO4, 1.00; K2HPO4, 0.61; MgSO4 × 7 H2O, 1.00; FeSO4 × 7 H2O, 0.0066; para-aminobenzoic acid, 0.10; and ammonium acetate, 2.90 and with either 50.0 glucose. Frozen samples were thawed on ice and RNA was isolated using high pure RNA isolation kit (Roche Diagnostics, Basel, Switzerland). Quality and concentration of RNA samples were checked using a nanodrop machine (Thermo Fisher Scientific, Waltham, Massachusetts, USA). The absence of DNA in the RNA samples was evaluated by qPCR analysis performed with BioRad CFX 96 Touch™. The RNA quality and integrity were determined using the Qsep 100 bioanalyzer (Bioptic Inc.). Messenger-RNA was depleted with the Ribo-Zero Magnetic Kit and a 250-300 bp insert cDNA library was constructed using the Illumina True seq stranded kit.
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-11-26T04:03:05Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-12-14T14:38:22Z
attributes.tag: External Id
attributes.value: SAMEA5176272
attributes.tag: INSDC center name
attributes.value: Wageningen Food & Biobased Research
attributes.tag: INSDC first public
attributes.value: 2019-11-26T04:03:05Z
attributes.tag: INSDC last update
attributes.value: 2018-12-14T14:38:22Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-7481:WT_3_4H
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: genotype
attributes.value: wild type genotype
attributes.tag: sample name
attributes.value: E-MTAB-7481:WT_3_4H
attributes.tag: scientific_name
attributes.value: Clostridium beijerinckii NCIMB 8052