Illumina HiSeq 2000 sequencing; ChIP-seq of Bacillus subtilis wild type and RNase J1 null strains
ChIP-seq of Bacillus subtilis wild type and RNase J1 null strains
Bacillus subtilis 168 wild-type (LK1371) and RNase J1 null strain (LK1381) were grown into exponential phase (OD600 ∼0,5) at 37°C in LB medium without any antibiotic. In RNase J1 null strain, the gene encoding RNase J1, is replaced by kanamycin antibiotic resistance Cells were crosslinked with 1% formaldehyde, stopped crosslink with 125mM glycin, washed and pelleted. Cells were sonicated to obtain 200-500bp long DNA. One miligram of total protein was bound into Dynabeads – antibody complex. Antibody against RNA polymerase β (8RB13, santacruz) was used. DNA was eluated, decrosslinked and purified with Nucleotide Removal Kit (QIAGEN). Strand specific libraries were prapared with NEBNext ChIP-Seq Library Prep Master Mix Set for Illumina (Biolabs) according to manufactuer´s instructions (Heidelberg, Germany).
Laboratory of Microbial Genetics  and Gene Expression, Institute of Microbiology of the CAS, v.v.i.
E-MTAB-5659:KOB_s