|
| Status |
Public on Mar 06, 2012 |
| Title |
prdm-control, biological rep1 |
| Sample type |
RNA |
| |
|
| Source name |
SV cells isolated from inguinal White adipose tissue
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6J
Sex: male
expression: sh-PRDM16
agent: none
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from cells or tissues using Trizol (Invitrogen).
|
| Label |
biotin
|
| Label protocol |
cDNA was prepared using a T7 promoter-tailed oligo-dT primer, followed by second strand synthesis. Double-stranded cDNA was used as the template for in vitro transcription (IVT) catalyzed by T7 polymerase and containing biotinylated CTP and UTP. Biotinylated complementary RNA (cRNA) is purified using the RNeasy system (Qiagen).
|
| |
|
| Hybridization protocol |
The purified cRNA was fragmented for hybridization and purified by phenol/chloroform extraction and ethanol precipitation. The fragmented cRNA and several biotinylated control oligonucleotides (for quality control) were hybridized to a mouse 430_2 chip overnight at 45C.
|
| Scan protocol |
Each cRNA bound at its complementary oligonucleotide is excited using a confocal laser scanner, and the positions and intensities of the fluorescent emissions are captured.
|
| Data processing |
Array normalization, expression value calculation and clustering analysis were performed using DNA-Chip Analyzer (dChip). The Invariant Set Normalization method was used to normalize arrays at probe cell level to make them comparable, and the model-based method was used for probe-selection and computing expression values.
|
| |
|
| Submission date |
Jan 11, 2012 |
| Last update date |
Mar 06, 2012 |
| Contact name |
Kosaku Shinoda |
| E-mail(s) |
[email protected]
|
| Phone |
415-476-9736
|
| Fax |
415-514-2346
|
| URL |
http://www.diabetes.ucsf.edu/members/shingo-kajimura
|
| Organization name |
University of California, San Francisco
|
| Department |
UCSF Diabetes Center and Department of Cell and Tissue Biology
|
| Street address |
35 Medical Center Way
|
| City |
San Francisco |
| ZIP/Postal code |
94143-0669 |
| Country |
USA |
| |
|
| Platform ID |
GPL8321 |
| Series (1) |
| GSE35011 |
PPARg agonists induce a white-to-brown fat conversion through stabilization of PRDM16 protein |
|
