|
| Status |
Public on Sep 14, 2013 |
| Title |
Ovaries, 3wks, SD (10hrs of Light) M43 |
| Sample type |
RNA |
| |
|
| Source name |
Ovaries, 3wks, SD (10hrs of Light)
|
| Organism |
Phodopus sungorus |
| Characteristics |
treatment: 10hrs of Light
tissue: Ovaries
Sex: female
age: 3wks
|
| Treatment protocol |
Female hamsters from both LD and SD groups were euthanized by CO2 Chamber at the age of 3 week or 8 week, respectively. Ovaries were asecptically removed,snap-frozen on dry ice, and stored at -80 degree until RNA extraction.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Both ovaries from each hamster were homogenized with a Polytron, and total RNA was extracted with Trizol (Invitrogen, Carlsbad, CA) according to the manufacturer's instructions. RNA was purified again with Rneasy Kit (Qiagen, Valencia, CA), examined its quantity and quality with and Agilent Bioanlyzer 2100 (Agilent Technologies,Palo Alto, CA).
|
| Label |
Cy3
|
| Label protocol |
One ug of total RNA is amplified and labeled with Cyanine-3 (Cy3) using Ambion Amino Allyl MessageAmp™ II aRNA Amplification Kit.
|
| |
|
| Hybridization protocol |
Cy3-labelled cRNA was hybridized to 4X44K microarray and washed, according to manufacturer's protocols. Briefly, Cy3-labelled cRNA was fragmented at 60°C for 30 minutes in a reaction volume of 55 ul containing 1x Agilent fragmentation buffer and 2x Agilent blocking agent following the manufacturers instructions. On completion of the fragmentation reaction, 55 ul of 2x Agilent hybridization buffer was added to the fragmentation mixture and hybridized to Agilent Mouse 44K custom high-definition whole-genome oligo array (G2519F-014868) for 17 hours at 65°C in a rotating Agilent hybridization oven. After hybridization, microarrays were washed 1 minute at room temperature with GE Wash Buffer 1 (Agilent) and 1 minute with 37°C GE Wash buffer 2 (Agilent), then dried immediately by brief centrifugation.
|
| Scan protocol |
Agilent microarray scanner was used to scan the microarry using one color scan setting.
|
| Description |
Gene expression in ovary from female hamster raised in SD(10 hours of light per day) by 3wks of age
|
| Data processing |
The Scanned images were analyzed with Agilent Feature Extraction software by obtaining fluorecent signal.
|
| |
|
| Submission date |
Sep 13, 2013 |
| Last update date |
Sep 14, 2013 |
| Contact name |
Ned Place |
| E-mail(s) |
njp27@cornell.edu
|
| Organization name |
Cornell University
|
| Department |
Population Medicine & Diagnostic Sciences
|
| Street address |
240 Farrier Road
|
| City |
Ithaca |
| State/province |
NY |
| ZIP/Postal code |
14853 |
| Country |
USA |
| |
|
| Platform ID |
GPL4134 |
| Series (1) |
| GSE50869 |
Different developmental trajectories of ovaries in Siberian hamsters raised in long and short day lengths |
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