|
| Status |
Public on Feb 28, 2015 |
| Title |
Brown adipocytes differentiation day 0 |
| Sample type |
RNA |
| |
|
| Source name |
Brown adipocytes during differentiation
|
| Organism |
Mus musculus |
| Characteristics |
cell type: brown adipocyte
genotype: wild type
feeding: N/A
|
| Treatment protocol |
Mice were injected intraperitoneally daily with saline or CL316,243 (1 mg/kg) for 7 days. Brown adipocytes were differentiated for 7 days.
BAT and WAT lysate were harvested after treatment. Brown adipocytes lysate were harvested during differentiation.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
Cyanine-3 (Cy3) labeled cRNA was prepared from 0.5 ug RNA using the One-Color Low RNA Input Linear Amplification PLUS kit (Agilent) according to the manufacturer's instructions, followed by RNAeasy column purification (QIAGEN, Valencia, CA). Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
1.5 ug of Cy3-labelled cRNA (specific activity >10.0 pmol Cy3/ug cRNA) was fragmented at 60°C for 30 minutes in a reaction volume of 250 ml containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 250 ml of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Whole Human Genome Oligo Microarrays (G4112A) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Slides were scanned immediately after washing on the Agilent DNA Microarray Scanner (G2505B) using one color scan setting for 1x44k array slides (Scan Area 61x21.6 mm, Scan resolution 10um, Dye channel is set to Green and Green PMT is set to 100%).
|
| Description |
Gene expression data from brown adipocytes during differentiation day 0
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 9.1 (Agilent) using default parameters (protocol GE1-v1_91 and Grid: 012391_D_20060331) to obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
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| |
|
| Submission date |
May 13, 2014 |
| Last update date |
Feb 28, 2015 |
| Contact name |
xuyun zhao |
| E-mail(s) |
[email protected]
|
| Organization name |
Life Sciences Institute
|
| Street address |
210 washtenaw ave
|
| City |
Ann Arbor |
| State/province |
Michigan |
| ZIP/Postal code |
48109 |
| Country |
USA |
| |
|
| Platform ID |
GPL15691 |
| Series (1) |
| GSE57643 |
LncRNAs expression profiling in adipose tissues and during brown adipocyte differentiation |
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