|
| Status |
Public on Nov 25, 2009 |
| Title |
Untreated replicate 3 |
| Sample type |
RNA |
| |
|
| Source name |
Spleen - Untreated
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
age: 8 weeks
tissue: Spleen
|
| Treatment protocol |
Spleen of 8 weeks old mice was isolated and B cells were purified and stimulated with IL-4 (Pepro-Tech) and CD40L (R&D Systems) as indicated at 5 ng/ml and 200 ng/ml, respectively. The BCR of stimulated cells was activated by incubation with goat F(ab’)2 anti-mouse IgM (Southern Biotechnologies) at 2.5 μg/ml for 3 hour.
|
| Growth protocol |
Mice were maintained for 8 weeks in a controlled environment with a 12 h dark/light cycle.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using Trizol Reagent (Invitrogen) and further purified using QIAGEN RNeasy mini kit in accordance with the prescribed protocol provided with the kit. Quality control was performed with Agilent Bioanalyser.
|
| Label |
Cy3
|
| Label protocol |
Total RNA was amplified and purified to yield biotinylated cRNA using an Ambion Illumina RNA amplification kit (Ambion).
|
| |
|
| Hybridization protocol |
Labeled cRNA (1.5 ug) were hybridized to the Sentrix MouseRef-6 Expression Beadchip for 16-18 h at 58 °C, according to the manufacturer's instructions (Illumina).
|
| Scan protocol |
Following attachment of Amersham fluorolink streptavidin-Cy3 (GE Healthcare Bio-Sciences), arrays were scanned with an Illumina bead array reader.
|
| Description |
control
|
| Data processing |
Signal intensities of genes were normalized by cubic spline method.
|
| |
|
| Submission date |
Apr 08, 2009 |
| Last update date |
Nov 25, 2009 |
| Contact name |
Thomas Grundström |
| E-mail(s) |
thomas.grundstrom@molbiol.umu.se
|
| Phone |
+46 90 785 2531
|
| Organization name |
Umeå University
|
| Department |
Department of Molecular Biology
|
| Lab |
TGN Lab
|
| Street address |
Analysvägen 1
|
| City |
Umeå |
| ZIP/Postal code |
901 87 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL6887 |
| Series (1) |
| GSE15606 |
Whole genome transcription profile of antigen receptor activated B cells |
|
