|
| Status |
Public on Nov 18, 2010 |
| Title |
Mouse pituitary gland, biological replicate 2 |
| Sample type |
RNA |
| |
|
| Source name |
Mouse pituitary gland
|
| Organism |
Mus musculus |
| Characteristics |
tissue: pituitary gland
strain: C57Bl6
Sex: male
age: 10-12 weeks old
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA from mouse tussues was extracted using Trizol reagent according to the manufacturer's protocol (Invitrogen), followed by a cleanup procedure with Rneasy colmuns (Qiagen). RNA from mouse islets as well as mouse pituitary and adrenal gland was extracted using the absolutely RNA microprep (Stratagene). For MIN6 cells we used the PureLink micro-to-midi RNA kit (INvitrogen)
|
| Label |
Biotin
|
| Label protocol |
Cellular mRNA was reverse transcribed into cDNA (SuperScript Choice System Invitrogen, Carlsbad, CA using oligo-dT primers and a T7 RNA polymerase promoter site). Two ug of total RNA was used to prepare biotinylated cRNA with IVT labeling kit (Affymetrix, Santa Clara, CA) according to the Genechip expression analysis technical manual 701025 Rev.5. The concentration of labelled cRNA was measured using the NanoDrop ND-1000 spectrophotometer. Labeled cRNA was fragmented in a fragmentation buffer during 35 min at 94C. The quality of labelled and fragmented cRNA was analysed using the Agilent bioanalyzer 2100.
|
| |
|
| Hybridization protocol |
Fragmented cRNA was hybridised to mouse 430 2.0 arrays (Affymetrix) during 16h at 45C.
|
| Scan protocol |
The arrays were washed and stained in a fluidics station (Affymetrix) and scanned using the Affymetrix 3000 GeneScanner.
|
| Description |
CEL file uploaded in GSE9954
GSM252097
|
| Data processing |
All images were analyzed using GCOS version 1.4 with the MAS5 algorithm
|
| |
|
| Submission date |
Sep 17, 2010 |
| Last update date |
Nov 18, 2010 |
| Contact name |
Katleen Lemaire |
| E-mail(s) |
[email protected]
|
| Organization name |
K.U.Leuven
|
| Street address |
Herestraat 49, bus 901
|
| City |
Leuven |
| ZIP/Postal code |
3000 |
| Country |
Belgium |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE24207 |
mRNA analysis in different mouse tissues |
|
| Relations |
| Reanalysis of |
GSM252097 |
