|
| Status |
Public on Feb 01, 2016 |
| Title |
striatum-R6/2 x DREAM +/- REP3 |
| Sample type |
RNA |
| |
|
| Source name |
striatum-R6/2 x DREAM +/-
|
| Organism |
Mus musculus |
| Characteristics |
strain: B6xCBA
genotype/variation: R6/2 x DREAM +/-
cell type: striatal neurons
|
| Growth protocol |
Preparation of transgenic mice expressing a dominant active mutant of DREAM has been reported before (Gomez-Villafuertes et al., 2005; Wu et al., 2010). DREAM knockout mice {Cheng, 2002 #95} were crossed with R6/2 mice to generate R6/2xDREAM+/- mice. The R6 mice were originally from Jackson laboratories.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA from whole striatum from wild type and transgenic mice was prepared using TRIzol (Invitrogen) and the RNAeasy Mini Kit (Qiagen). RNA was quantified and the quality was assessed with a 2100 Bioanalyzer (Agilent technologies).
|
| Label |
biotin
|
| Label protocol |
cDNA was synthesized from 4 µg of total RNA using one-cycle target labeling and control reagents (Affymetrix) to produce biotin labeled cRNA. The cRNA preparation (15 µg) was fragmented at 94ºC for 35 min into 35-200 bases in length.
|
| |
|
| Hybridization protocol |
Labeled cRNAs were hybridized to Affymetrix chips (GeneChip Mouse Genome 430 2.0 Array). Each sample was added to a hybridization solution containing 100 mM 2-(N-morpholino) ethanesulfonic acid, 1 M Na+, and 20 mM of EDTA in the presence of 0.01% of Tween-20 to a final cRNA concentration of 0.05 µg/ml. Hybridization was performed for 16 h at 45ºC. Each microarray was washed and stained with streptavidin-phycoerythrin in a Fluidics station 450 (Affymetrix)
|
| Scan protocol |
GeneChips were scanned at 1.56 µm resolution in a GeneChip® Scanner 3000 7G System (Affymetrix)
|
| Description |
KR3
|
| Data processing |
GeneChip intensities were background-corrected, normalized and sumarized by RMA method, using the “Affy” package from Bioconductor.
|
| |
|
| Submission date |
Jun 19, 2013 |
| Last update date |
Feb 01, 2016 |
| Contact name |
Juan Carlos Oliveros |
| Organization name |
CNB, CSIC
|
| Street address |
Darwin 3
|
| City |
Cantoblanco |
| State/province |
Madrid |
| ZIP/Postal code |
28049 |
| Country |
Spain |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE48104 |
DREAM silencing is part of a neuroprotective mechanism in Huntington's disease |
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