Sample 2
Bacillus subtilis subsp. subtilis str. 168
Protocols: Bacillus subtilis wt, ∆sigI-rsgI and ∆rsgI strains were inoculated from single colony to 10 ml of LB medium. Cultures were grown overnight at 37°C. Next day, the cultures were inoculated into 20 ml of LB medium (RT) to OD600∼0.01 and grown at 37°C and 52°C, respectively. Cells were harvested in exponential phase (OD600∼0.5). Three ml of culture were immediately treated with a double volume of RNAprotect Bacteria reagent (QIAGEN) for 5 min at RT to prevent degradation of RNA. Cells were pelleted and frozen immediately. In all steps of cultivation (with the exception of the last step of cultivation for total RNA isolation) medium for ∆sigI-rsgI strain was supplemented with spectinomycin 100 μg/ml; medium for ∆rsgI strain was supplemented with linkomycin 12.5 μg/ml and erythromycin 0.5 μg/ml. The experiment was repeated three times. RNA was extracted using RNeasy Mini Kit 50 (QIAGEN) according to the manufacturer's protocol. Finally, RNA was treated twice with DNase (TURBO DNA-free Kit, Ambion). Two micrograms of total RNA were rRNA-depleted with RiboMinus Transcriptome Isolation Kit, bacteria (Invitrogen). A strand-specific library was prepared for each sample with Illumina compatible NEXTflex Rapid Directional RNA-Seq Kit (Bioo Scientific) according to manufacturer's protocol.
ENA first public
2018-05-31
ENA last update
2017-12-15
External Id
SAMEA104431009
INSDC center alias
Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology, v.v.i. AS CR
INSDC center name
Laboratory of Microbial Genetics and Gene Expression, Institute of Microbiology, v.v.i. AS CR
INSDC first public
2018-05-31T17:01:57Z
INSDC last update
2017-12-15T11:03:28Z
INSDC status
public
Submitter Id
E-MTAB-6314:Sample 2
broker name
ArrayExpress
genetic modification
deletion of the whole operon
genotype
sigI-rsgI knockout
growth condition
37C LB broth
replicate
1
sample name
E-MTAB-6314:Sample 2
strain
168
type-material
type strain of Bacillus subtilis