|
| Status |
Public on May 01, 2010 |
| Title |
MOH2wt Sm_infected 01 |
| Sample type |
RNA |
| |
|
| Source name |
Cecum, WT, infected, 10h
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57Bl/6J
tissue: cecum
genotype: wildtype
treatment: Salmonella SB300 infection
infection duration: 10h
|
| Treatment protocol |
Streptomycin-pretreated mice (20 mg/animal) were infected by gavage (5 x 10E7 cfu) as published previously. Live bacterial loads in cecal content were determined by plating. Experiments were approved and performed as legally required (Licence 201/2007).
|
| Growth protocol |
Wildtype and isogenic PARP1 knockout mice were bred in a C57Bl/6J background. Regular back-crossings were performed to maintain isogenicity. All mice were bred and kept in a specified pathogen-free area.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
The intestinal tract was excised and the cecum was isolated. The cecum was then cut into four slices. Slices 1 and 3 were washed in cold PBS to remove cecum content and afterwards frozen in liquid nitrogen in 300 μl RLT-buffer (RNeasy Mini Kit, Qiagen) with 1% β-Mercaptoethanol and stored at -80°C till RNA-extraction. Frozen tissue was homogenized in RLT for 3 minutes at 25 Hz using a Tissue Lyser (Qiagen). RNA was processed using the RNeasy Mini Kit (Qiagen) and the RNase-Free DNase Set (Qiagen) according to the manufacturer’s instruction.
|
| Label |
Cy3
|
| Label protocol |
1.6 - 5 μg of total RNA were reverse transcribed to cDNA, and amplified and labeled to cRNA with the Agilent Low RNA Input Linear Amplification Kit PLUS (order number 5184-3525, Agilent).
|
| |
|
| Hybridization protocol |
Agilent Gene Expression kit for oligo microarrays (Agilent, order number 5188-5242, protocol G4140-90050).
|
| Scan protocol |
Agilent Feature Extraction software 9.5.3.1.
|
| Description |
MOH2wt Sm_infected 01
|
| Data processing |
GeneSpring GX10 default loading and analysis.
|
| |
|
| Submission date |
Nov 24, 2009 |
| Last update date |
Nov 24, 2009 |
| Contact name |
Hubert Rehrauer |
| E-mail(s) |
[email protected]
|
| Organization name |
ETH Zurich / University of Zurich
|
| Department |
FGCZ
|
| Street address |
Winterthurerstr. 190
|
| City |
Zurich |
| ZIP/Postal code |
8057 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL4134 |
| Series (1) |
|
