title: Illumina HiSeq 2000 sequencing; RNA-seq of Bacillus subtilis wild type and RNase J1 null strains
design: RNA-seq of Bacillus subtilis wild type and RNase J1 null strains
library_construction_protocol: Bacillus subtilis 168 wild-type (LK1371) and RNase J1 null strain strains (LK1381) were grown at 37°C in LB medium and harvested in exponential phase (OD600 ∼0,5). In RNase J1 null strain, the gene encoding RNase J1 is replaced by kanamycin antibiotic resistance. Two ml exponentially growing cells were treated with RNAprotect Bacteria reagent (QIAGEN), pelleted and immediately frozen. RNA was isolated by RNas Mini Kit (QIAGEN). Finally, RNA was DNase treated (TURBO DNA-free Kit, Ambion). Five micrograms of total RNA were rRNA depleted with Ribo-Zero Magnetic Kit, Gram-positive bacteria (Epicentre) and strand specific libraries prepared with Illumina compatible NEXTflex Rapid Directional RNA-Seq Kit (Bioo Scientific) according to manufactuer´s instructions
attributes.tag: Experimental Factor: genotype
attributes.value: RNase J1 deletion