|
Status |
Public on Feb 28, 2017 |
Title |
PZN_Sterne2 |
Sample type |
SRA |
|
|
Source name |
Bacterial cells, plantazolicin
|
Organism |
Bacillus anthracis str. Sterne |
Characteristics |
strain/background: Sterne treatment compound: plantazolicin
|
Treatment protocol |
Samples were treated with 0.25X MIC compound (plantazolicin or Me2-Arg-Az5) or an equivalent volume of DMSO, and allowed to incubate for 10 min at 37 °C. Samples were treated with Qiagen RNAprotect bacterial reagent and harvested at 4000 x g for 10 min.
|
Growth protocol |
(3) independent 3 mL cultures for each treatment group were grown to optical density (600 nm) of 0.4.
|
Extracted molecule |
total RNA |
Extraction protocol |
Harvested cells were resuspended in 250 μl of 10 mM Tris (pH 8.5) with 15 mg/ml lysozyme and 5 μl proteinase K (20 mg/ml) and digested for 1 h at 22°C with gentle agitation. RNA was extracted using the Qiagen RNeasy Mini Kit. An on-column DNase I digestion was performed using the Qiagen RNase-free DNase set according to manufacturer's instructions. RNA isolation protocol was completed according to manufacturer's instructions, followed by a second DNase I digestion and an RNA cleanup protocol. Libraries were created using the TruSeq Stranded RNA Sample Prep kit after rRNA depletion using the RiboZero Bacteria kit, following manufacturer's instructions.
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|
|
Library strategy |
RNA-Seq |
Library source |
transcriptomic |
Library selection |
cDNA |
Instrument model |
Illumina HiSeq 2000 |
|
|
Description |
Bacillus anthracis Sterne 7702 mRNA Processed data file: NC_005945_transcripts_DMSO_PZN.txt Processed data file: NC_005945_transcripts_PZN_Az5.txt
|
Data processing |
Data was processed with the Rockhopper version 1.30 pipeline using B. anthracis Sterne (NC_005945.1) and B. anthracis Ames Ancestor plasmid pXO1 (NC_007322.2) as references. Default values (allowed mismatches 0.15, minimum seed length 0.33, minimum expression of UTRs and ncRNAs 0.5) were used, with the exception that reverse complement reads were used for mapping. Genome_build: B. anthracis Sterne: NC_005945.1 Genome_build: B. anthracis Ames Ancestor plasmid pXO1: NC_007322.2 Supplementary_files_format_and_content: Tab-delimited text files include expression values that are counts normalized using the Rockhopper pipeline.
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|
|
Submission date |
Sep 22, 2015 |
Last update date |
May 15, 2019 |
Contact name |
Douglas A Mitchell |
E-mail(s) |
douglasm@illinois.edu
|
Phone |
2173331345
|
Organization name |
University of Illinois at Urbana-Champaign
|
Department |
Chemistry
|
Lab |
Mitchell Lab
|
Street address |
1206 W. Gregory Dr. MC-195 Room 3105 IGB
|
City |
Urbana |
State/province |
IL |
ZIP/Postal code |
61801 |
Country |
USA |
|
|
Platform ID |
GPL20947 |
Series (1) |
GSE73343 |
Bacillus anthracis gene expression upon plantazolicin and Me2-Arg-Az5 |
|
Relations |
BioSample |
SAMN04102795 |
SRA |
SRX1275047 |