|
Status |
Public on Feb 03, 2015 |
Title |
R.sphaeroides 2.4.1 ∆iscR vs. R.sphaeroides 2.4.1 wildtype replicate 2 |
Sample type |
RNA |
|
|
Channel 1 |
Source name |
2.4.1∆iscR
|
Organism |
Cereibacter sphaeroides 2.4.1 |
Characteristics |
genotype: ∆iscR strain: 2.4.1
|
Growth protocol |
R. sphaeroides strains grown microaerobically
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated by using the hot phenol method (Janzon et al., 1986). After DNase I treatment the RNA was purified by using mixtures of phenol-chloroform-isoamylalcohol and chloroform-isoamylacohol. For microarray analysis the RNA was further purified by RNeasy®MinElute™ spin columns (Qiagen) following the manufacturer’s instructions.
|
Label |
Cy3
|
Label protocol |
The ULSTM Fluorescent Labeling Kit for Agilent arrays (Kreatech) was used for RNA labeling and fragmentation
|
|
|
Channel 2 |
Source name |
WT 2.4.1
|
Organism |
Cereibacter sphaeroides 2.4.1 |
Characteristics |
genotype: wild type strain: 2.4.1
|
Growth protocol |
R. sphaeroides strains grown microaerobically
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was isolated by using the hot phenol method (Janzon et al., 1986). After DNase I treatment the RNA was purified by using mixtures of phenol-chloroform-isoamylalcohol and chloroform-isoamylacohol. For microarray analysis the RNA was further purified by RNeasy®MinElute™ spin columns (Qiagen) following the manufacturer’s instructions.
|
Label |
Cy5
|
Label protocol |
The ULSTM Fluorescent Labeling Kit for Agilent arrays (Kreatech) was used for RNA labeling and fragmentation
|
|
|
|
Hybridization protocol |
Total RNA of three independent experiments of an iscR deletion strain (2.4.1∆iscR) and a control strain (wild type 2.4.1) were pooled and hybridized to one array. Two arrays werehybridized with independent experimental runs. Gene chip hybridization and scanning were performed according to the specifications from Agilent.
|
Scan protocol |
Scanned on an Agilent High Resolution Microarray Scanner. Images were quantified using Agilent Feature Extraction Software (version A.7.5)
|
Description |
biological sample 4-6
|
Data processing |
R with Limma package was used for data evaluation. LOESS normalized and background subtracted.
|
|
|
Submission date |
Feb 02, 2015 |
Last update date |
Feb 03, 2015 |
Contact name |
Gabriele Klug |
E-mail(s) |
Gabriele.Klug@mikro.bio.uni-giessen.de
|
Organization name |
Justus-Liebig University Gießen
|
Department |
Molecular- and Microbiology
|
Street address |
Heinrich-Buff-Ring 26-32
|
City |
Gießen |
ZIP/Postal code |
35392 |
Country |
Germany |
|
|
Platform ID |
GPL15457 |
Series (1) |
GSE65537 |
IscR of Rhodobacter sphaeroides functions as repressor of genes for iron-sulphur metabolism and represents a new type of iron-sulphur binding protein |
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