|
| Status |
Public on Oct 19, 2010 |
| Title |
BAT from per2-/- mice, biological rep1 |
| Sample type |
RNA |
| |
|
| Source name |
BAT from 20-week-old per2-/- mouse
|
| Organism |
Mus musculus |
| Characteristics |
genotype: per2-/-
gender: male
age: 20 weeks
tissue: fresh brown adipose tissue
|
| Treatment protocol |
All mice were fed a chow diet (ProLab RMH2500 5P14) ad libitum.
|
| Growth protocol |
Per2-/- and per2+/+ mice (Bae et al., 2001) were grown under 12 hr light/12 hr dark (LD) cycles.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was initially isolated using TRIzol Reagent (Gibco BRL Life Technologies, Rockville, MD), and passed through an RNeasy spin column (Qiagen, Chatsworth, CA) for further clean up. Eluted total RNA was quantified (Nanodrop 1000) with a portion of the recovered total RNA adjusted to a final concentration of 250ng/ul.
|
| Label |
biotin
|
| Label protocol |
The fragmented single-stranded DNA was end-labeled with recombinant terminal deoxynucleotidyl transferase (TdT) and the Affymetrix proprietary DNA Labeling Reagent that is covalently linked to biotin.
|
| |
|
| Hybridization protocol |
Following the recommended procedure, 0.54 ug of this fragmented target single-stranded cDNA was hybridized at 45°C with rotation for 17 hours (Affymetrix GeneChip® Hybridization Oven 640) to probe sets present on an Affymetrix array. The GeneChip® arrays were washed and then stained (SAPE, streptavidin-phycoerythrin) on an Affymetrix Fluidics Station 450 (Fluidics protocol FS450_007).
|
| Scan protocol |
Arrays were scanned using the GeneChip Scanner 3000 7G and the cell intensities (*.DAT and *.CEL files) were obtained using Affymetrix’s GeneChip Operating Software, Ver. 1.4 (GCOS).
|
| Description |
Gene expression data from BAT of 20-week-old per2-/- mouse.
|
| Data processing |
The PLIER (Probe Logarithmic Intensity ERror) algorithm developed by Affymetrix was used.
Data were analyzed with Cyber-T software (http://cybert.microarray.ics.uci.edu/).
|
| |
|
| Submission date |
Feb 02, 2010 |
| Last update date |
Oct 19, 2010 |
| Contact name |
benedetto grimaldi |
| E-mail(s) |
bgrimald@uci.edu
|
| Phone |
949 824 8056
|
| Organization name |
university of california, irvine
|
| Department |
pharmacology
|
| Lab |
Sassone-Corsi
|
| Street address |
2226B Gillespie NRF
|
| City |
Irvine |
| State/province |
CA |
| ZIP/Postal code |
92697 |
| Country |
USA |
| |
|
| Platform ID |
GPL6246 |
| Series (1) |
| GSE20165 |
Expression data from white and brown adipose tissue (WAT and BAT) of per2-/- and control mice |
|
