|
| Status |
Public on Jan 12, 2010 |
| Title |
Pb7F_rep2_Array3 |
| Sample type |
RNA |
| |
|
| Source name |
whole body, 7-day-old adult, female, replicate 2
|
| Organism |
Drosophila pseudoobscura bogotana |
| Characteristics |
tissue: whole body
gender: Female
age: 7-day-old
|
| Growth protocol |
flies were reared on cornmeal/yeast medium and maintained at 20°C with a 12:12-h light/dark cycle.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
flies were instantly frozen in liquid nitrogen and stored in TRIZOL (Invitrogen) at -80°C. RNA was isolated using TRIZOL from at least 40 individuals. RNA quantity and quality were separately measured on a NanoDrop spectrophotometer (Thermo Scientific) and a Bioanalyzer 2100 (Agilent Technologies). High-quality samples (260/280>1.85; 260/230>1.7) from all lines of each species were pooled in equal amounts and further purified using the RNeasy kit (Qiagen).
|
| Label |
Cy3
|
| Label protocol |
Fluorescent cRNA was synthesized from 200 ng of total RNA using Agilent’s low-RNA input fluorescent linear amplification kit following manufacturer's protocols (Agilent Technologies). Labelled RNAs were cleaned with RNeasy columns (Qiagen), and cRNA yields were quantified on a NanoDrop spectrophotometer. Two separate labeling reactions per sample were pooled and hybridized to the arrays following Agilent’s protocols.
|
| |
|
| Hybridization protocol |
Agilent’s protocols
|
| Scan protocol |
Agilent’s protocols
|
| Description |
Global gene expression of 7-day-old virgin adults
|
| Data processing |
Performed in HDBStat: data is normalized by flagging out bad spots, filtering unexpressed genes (average raw intensity across all the samples <50), negative and positive Agilent controls, and then normalized with Chip Mean and log2 transformation.
|
| |
|
| Submission date |
Jul 20, 2009 |
| Last update date |
Jan 12, 2010 |
| Contact name |
Carlos Alberto Machado |
| E-mail(s) |
zjiang@umd.edu
|
| Organization name |
University of Maryland
|
| Department |
department of biology
|
| Street address |
1210 BPS
|
| City |
college park |
| State/province |
MD |
| ZIP/Postal code |
20742 |
| Country |
USA |
| |
|
| Platform ID |
GPL8884 |
| Series (1) |
| GSE17192 |
Evolution of sex-dependent gene expression in three recently diverged species of Drosophila |
|
