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Status |
Public on Jul 07, 2014 |
Title |
Wnt3a treated DRG culture rep1 |
Sample type |
RNA |
|
|
Source name |
mouse DRG tissue
|
Organism |
Mus musculus |
Characteristics |
strain: C57Bl/6 age: 4-6 weeks tissue: neuronal-enriched DRG culture treatment: Wnt3a
|
Treatment protocol |
The DRG cultures were serum-starved for 1h and then treated with Wnt3a (200 ng/ml) or vehicle (PBS) for 6h.
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Growth protocol |
DRGs from 6-8 weeks old mice were collected in cold PBS after removing axons and meningeal tissue, and digested for 30 min at 37°C in trypsin, collagenase and DNAse. Neurons were than separated from non-neuronal cells using a discontinuous Percoll gradient, plated on poly-L-lysine coated coverslips and cultured in F12 medium plus 10% serum in presence of a mixture of growth factors (BDNF 10 ng/ml, NGF 10 ng/ml, GDNF 5 ng/ml, NT3 5ng/ml, all from SIGMA) and mitotic inhibitor AraC 5 µM (SIGMA) for 3 days before treatment.
|
Extracted molecule |
total RNA |
Extraction protocol |
Total RNA was extracted with Trizol (Life Technologies) followed by clean-up and DNase I treatment with Qiagen RNeasy mini kit in accordance with the prescribed protocol provided with the kit. RNA quality control was performed with Agilent Bioanalyzer.
|
Label |
biotin
|
Label protocol |
Biotinylated cRNA was prepared with the Ambion TotalPrep kit for Illumina arrays.
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|
|
Hybridization protocol |
Microarrays were hybridized following Illumina's standard hybridization protocol on MouseRef-8 v2.0 expression BeadChips.
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Scan protocol |
Processed arrays were scanned on Illumina iScan System array reader according to the manufacturer's instructions.
|
Description |
replicate 1
|
Data processing |
Raw array data were processed with Illumina BeadStudio software (v3.2) using background subtraction and cubic-spline normalisation.
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Submission date |
Apr 17, 2014 |
Last update date |
Jul 07, 2014 |
Contact name |
Emil Karaulanov |
Organization name |
Institute of Molecular Biology
|
Lab |
Bioinformatics Core Facility
|
Street address |
Ackermannweg 4
|
City |
Mainz |
ZIP/Postal code |
55128 |
Country |
Germany |
|
|
Platform ID |
GPL6885 |
Series (1) |
GSE56880 |
Genome-wide transcriptome analysis of neuron-enriched DRG cultures exposed to Wnt3a |
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