|
| Status |
Public on Jan 31, 2018 |
| Title |
Adipose.14d.HFD.rep3 |
| Sample type |
RNA |
| |
|
| Source name |
adipose tissue
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57Bl/6
diet: 14 day high fat
|
| Treatment protocol |
Mice were fed a standard chow (13% of calories from fat) diet until 6 weeks of age and then either continued on standard chow, or fed a high fat diet (HFD, 46% of calories from fat) for 5 days or 14 days.
|
| Growth protocol |
C57Bl/6 mice were group housed on a 12-h light/dark cycle with free access to food and water.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using Qiagen Kit.
|
| Label |
biotin
|
| Label protocol |
The methods for preparation of labeled cRNA and array hybridization were provided by Affymetrix (Santa Clara, CA). Briefly, 3 ug of total RNA was converted into double-stranded cDNA using a cDNA synthesis kit SuperScript Choice (Invitrogen, Carlsbad, CA) and a T7-(dT)24 oligomer primer (Integrated DNA Technologies, Coralville, IO). Double-stranded cDNA was purified using affinity resin Sample Cleanup Module Kit (Affymetrix). Labeled cRNA was generated using a T7 RNA polymerase and biotin-labeled nucleotides in an in vitro transcription reaction (Enzo Diagnostics, Farmingdale, NY). The labeled cRNA was purified using Sample Cleanup Module Kit (Affymetrix).
|
| |
|
| Hybridization protocol |
Fifteen micrograms of labeled cRNA was fragmented by incubating at 94°C for 30 min in 40 mM Tris-acetate pH 8.1, 100 mM potassium acetate and 30 mM magnesium acetate. Ten micrograms of fragmented cRNA was then hybridized to Mouse Genome 430 2.0 arrays at 45°C for 19 hours in a rotisserie oven set at 60 rpm.
|
| Scan protocol |
Arrays were washed and stained in the Affymetrix Fluidics station and scanned on GeneChip scanner 3000.
|
| Data processing |
Data analysis files were generated using manufacturer’s GeneChip Command Console system (Affymterix).
|
| |
|
| Submission date |
Oct 11, 2016 |
| Last update date |
Jan 31, 2018 |
| Contact name |
Rima Chaudhuri |
| E-mail(s) |
rima.chaudhuri@sydney.edu.au
|
| Organization name |
University of Sydney
|
| Department |
Charles Perkins Center
|
| Street address |
D17 Johns Hopkins Drive
|
| City |
Camperdown, Sydney |
| State/province |
NSW |
| ZIP/Postal code |
2006 |
| Country |
Australia |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE87854 |
Insulin resistance in high fat diet mouse (adipose) |
|
