|
| Status |
Public on Apr 15, 2014 |
| Title |
Sleep Deprived Replicate 5 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
fly head, sleep deprived, 7 hours
|
| Organism |
Drosophila melanogaster |
| Characteristics |
light conditions: dark;dark
genotype: cyc01
age: 7 d
gender: Female
tissue: head
|
| Treatment protocol |
On the third day, they were either sleep deprived for 7 h according to standard procedures or starved for 7 h by replacing their food with 1% agar.
|
| Growth protocol |
Three day old female cyc01 mutants were monitored under baseline conditions for 2 days in dark;dark.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
Three milligrams of poly(A) mRNA was used as a template for each of cDNA synthesis reactions using Superscript II (GibcoBRL) in the presence of fluorescently labeled Cy3- or Cy5-dCTP (Amersham).
|
| |
|
| Channel 2 |
| Source name |
fly head
|
| Organism |
Drosophila melanogaster |
| Characteristics |
light conditions: dark;dark
genotype: cyc01
age: 7 d
gender: Female
tissue: head
|
| Treatment protocol |
On the third day, they were either sleep deprived for 7 h according to standard procedures or starved for 7 h by replacing their food with 1% agar.
|
| Growth protocol |
Three day old female cyc01 mutants were monitored under baseline conditions for 2 days in dark;dark.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
Three milligrams of poly(A) mRNA was used as a template for each of cDNA synthesis reactions using Superscript II (GibcoBRL) in the presence of fluorescently labeled Cy3- or Cy5-dCTP (Amersham).
|
| |
|
| |
| Hybridization protocol |
Reactions were purified using QIAQuick PCR purification columns (Qiagen), and labeled RNA was hybridized to clones arrayed on a 2x2-cm field under a plastic HybriSlip (Grace BioLabs) in a humidified cassette (TeleChem) for 15 h at 65 degrees C
|
| Scan protocol |
After the arrays were washed, fluorescence intensities were measured on an Agilent scanner.
|
| Description |
n/a
|
| Data processing |
Data from the 4,659 ESTs present were background subtracted, mean normalized and subjected to loess transformation using Standardization and Normalization of MicroArray Data (http://pevsnerlab.kennedykrieger.org/snomadinput.html). The two most extreme values were identified from each condition (control, sleep deprived and starved) and removed.
|
| |
|
| Submission date |
Oct 14, 2009 |
| Last update date |
Apr 15, 2014 |
| Contact name |
Paul Shaw |
| E-mail(s) |
shawp@pcg.wustl.edu
|
| Organization name |
Washington University School of Medicine
|
| Department |
Anatomy
|
| Lab |
Paul Shaw
|
| Street address |
660 S Euclid
|
| City |
St Louis |
| State/province |
MO |
| ZIP/Postal code |
63110 |
| Country |
USA |
| |
|
| Platform ID |
GPL9444 |
| Series (1) |
| GSE18550 |
Identification of Sleep Regulatory Genes in Flies Mutant for the Clock Gene cycle |
|
