|
| Status |
Public on Dec 16, 2006 |
| Title |
NPAS2 mutant replicate F20 |
| Sample type |
RNA |
| |
|
| Source name |
Aorta, male NPAS2 mutant, ZT14
|
| Organism |
Mus musculus |
| Characteristics |
Strain: NPAS2 mutant on C57Bl/6J background
Sex: male
Tissue: aorta
Circadian time: ZT14
|
| Treatment protocol |
Mice were initially acclimatized for at least 2 weeks to a 12 hour light/dark cycle (light 7:00-19:00h or ZT0-ZT12; dark 19:00-7:00h or ZT12-ZT24). Mice were placed in constant darkness for at least 30 hours before sacrifice.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Mice were anesthetized at the time of sacrifice. The abdomen and thorax were opened and a blood sample was taken from the vena cava. The animal was then perfused via the left ventricle with 10mls of phosphate buffered saline (PBS) (Fisher Scientific) followed by 5mls of DEPC treated water (Fischer Scientific). The aorta was exposed as close to the heart as possible at the aortic arch and to just beyond the femoral artery bifurcation, using a dissection microscope. It was then removed and immediately flash-frozen in liquid Nitrogen. Aortae were pulverized on dry ice with a Plattner’s diamond mortar and pestle set (Fisher Scientific). Total RNA was extracted using RNAqueous (Ambion, TX) and Trizol (Invitrogen, CA) and quantified by spectrophotometer.
|
| Label |
Biotin
|
| |
|
| Hybridization protocol |
Target cRNA was prepared from total RNA and hybridized using 20ug of cRNA onto Affymetrix MOE 430V2 chips in accordance with protocols described in the Affymetrix Gene-Chip Expression Analysis Technical Manual.
|
| Description |
After 2 week acclimation to 12 hr/12 hr light/dark regimen, mice were placed in constant darkness for at least 30 hours before sacrifice under anesthesia at ZT14. Thoracic aortae were harvested and the RNA extracted and hybridized to Affymetrix MOE 430V2 chips.
|
| Data processing |
GCRMA normalization; Affymetrix Microarray Suite 5.0 normalization
|
| |
|
| Submission date |
Dec 16, 2005 |
| Last update date |
Dec 19, 2005 |
| Contact name |
Radu Daniel Rudic |
| E-mail(s) |
rudicrd@spirit.gcrc.upenn.edu
|
| Phone |
215 898-0255
|
| Fax |
215 573-9004
|
| Organization name |
University of Pennsylvania
|
| Department |
Center of Experimental Therapeutics
|
| Lab |
Garret FitzGerald Lab
|
| Street address |
421 Curie Boulevard
|
| City |
Philadelphia |
| State/province |
PA |
| ZIP/Postal code |
19104 |
| Country |
USA |
| |
|
| Platform ID |
GPL1261 |
| Series (1) |
| GSE3849 |
Differential transcript abundance in NPAS2 mutant mouse at ZT14 |
|
