|
Status |
Public on Jan 25, 2016 |
Title |
pBAD-ctr.2 (control) |
Sample type |
mixed |
|
|
Channel 1 |
Source name |
Salmonella enterica SL1344_pBAD-ctr_genomic DNA
|
Organism |
Salmonella enterica subsp. enterica serovar Typhimurium str. SL1344 |
Characteristics |
genotype/variation: carrying pBAD-ctr treated with: 0.2% (final conc.) L-arabinose time point: 10 min post treatment
|
Treatment protocol |
At OD=1.5, 0.2% (final conc.) L-arabinose was added. Total RNA was isolated 10 min post treatment.
|
Growth protocol |
Salmonella carrying pBAD-ctr or pBAD-SdsR were grown in LB-media to OD=1.5.
|
Extracted molecule |
genomic DNA |
Extraction protocol |
SV40 RNA isolation Kit (promega). GenomicDNA was extracted using the Qiagen 'Genomic DNA' Kit
|
Label |
Cy5
|
Label protocol |
total RNA was labelled with Cy3 using random hexamers and reverse transcriptase (Stratagene AffinityScript) while genomicDNA was labelled with Cy5 using the Invitrogen BioPrime DNA Labeling System
|
|
|
Channel 2 |
Source name |
Salmonella enterica SL1344_pBAD-ctr_total RNA
|
Organism |
Salmonella enterica subsp. enterica serovar Typhimurium str. SL1344 |
Characteristics |
genotype/variation: carrying pBAD-SdsR treated with: 0.2% (final conc.) L-arabinose time point: 10 min post treatment
|
Treatment protocol |
At OD=1.5, 0.2% (final conc.) L-arabinose was added. Total RNA was isolated 10 min post treatment.
|
Growth protocol |
Salmonella carrying pBAD-ctr or pBAD-SdsR were grown in LB-media to OD=1.5.
|
Extracted molecule |
total RNA |
Extraction protocol |
SV40 RNA isolation Kit (promega). GenomicDNA was extracted using the Qiagen 'Genomic DNA' Kit
|
Label |
Cy3
|
Label protocol |
total RNA was labelled with Cy3 using random hexamers and reverse transcriptase (Stratagene AffinityScript) while genomicDNA was labelled with Cy5 using the Invitrogen BioPrime DNA Labeling System
|
|
|
|
Hybridization protocol |
Microarray slide were hybridized in a water bath over night at 63C. For detailed protocol see: http://www.ifr.ac.uk/safety/Microarrays/default.html#protocols
|
Scan protocol |
Axon GenePix4000A, 10nm resolution scan
|
Data processing |
Data centring was performed by bringing the median ratio (Cy5/Cy3) for each of the 16 blocks to one using the following equation: (Ti) = (Cy5i/Cy3i) - c, where T is the centred ratio, i is the gene index, Cy5 and Cy3 are the red and green intensities and c is the 50th percentile of all Cy5/Cy3 ratios. pBAD-ctr serves as control for pBAD-SdsR.
|
|
|
Submission date |
Jan 24, 2016 |
Last update date |
Jan 25, 2016 |
Contact name |
Kai Papenfort |
Organization name |
IMIB Würzburg
|
Department |
RNA Biology
|
Street address |
Josef-Schneider Str.2/BAU D15
|
City |
Würzburg |
State/province |
Würzburg |
ZIP/Postal code |
97080 |
Country |
Germany |
|
|
Platform ID |
GPL5780 |
Series (1) |
GSE77157 |
The target spectrum of SdsR small RNA in Salmonella |
|