|
| Status |
Public on Sep 16, 2004 |
| Title |
Rat fatty liver13 |
| Sample type |
RNA |
| |
|
| Source name |
Wistar rat liver tissue following animal exposure to orotic acid for 14 days
|
| Organism |
Rattus norvegicus |
| Characteristics |
Kyoto rat liver tissue following animal exposure to orotic acid for 1 day
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by RNA Isolation Kit (Stratagene) from the livers of Wistar and Kyoto rats at day 0 (Wistar n=3, Kyoto n=2), day 1 and 3 (Wistar n=2, Kyoto n=2), and day 14 (Wistar n=3, Kyoto n=2).
|
| Label |
biotin
|
| Label protocol |
cDNA was generated from total RNA using SuperscriptTM Double-stranded cDNA Synthesis kit (Invitrogen, Life Technologies), cRNA was prepared by in vitro transcription (BioArray High Yield RNA transcript labelling kit Affymetrix, UK), and purified on an affinity resin column (RNeasy, Qiagen Ltd., UK).
|
| |
|
| Hybridization protocol |
Biotinylated cRNA was fragmented and hybridised to RGU34A Affymetrix GeneChips according to standard Affymetrix protocols.
|
| Scan protocol |
Performed according to standard Affymetrix protocols.
|
| Description |
Rats (Charles River UK Ltd.) were fed either standard laboratory chow, or chow supplemented with 1% orotic acid (Sigma Aldrich, UK) ad libitum. Orotic acid feeding is a well-characterised method for inducing reversible fatty liver disease in rodents. Rats were killed by cervical dislocation and the left lateral lobe of the liver excised. Tissues were snap frozen and stored in liquid nitrogen and/or at –80oC.
Keywords = orotic acid
Lot batch = 1
|
| Data processing |
Data were analysed with the Affymetrix Microarray Suite 5.0 algorithm to generate a “signal” value and a “detection” label. Prior to normalisation, probes that generated an absent call in all cells were removed. Normalisation was calculated using a Global Scaling method based on the entire data set. Globally scaled values <10 were rounded up to 10.
|
| |
|
| Submission date |
Sep 15, 2003 |
| Last update date |
Jan 31, 2007 |
| Contact name |
Julian Griffin |
| E-mail(s) |
jlg40@mole.bio.cam.ac.uk
|
| Phone |
+44(0)1223 333626
|
| Organization name |
University of Cambridge
|
| Department |
Biochemistry
|
| Lab |
Griffin
|
| Street address |
Tennis Court Road
|
| City |
Cambridge |
| ZIP/Postal code |
CB2 1QW |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL85 |
| Series (1) |
| GSE665 |
Orotic acid induced fatty liver disease |
|
