|
| Status |
Public on Apr 08, 2012 |
| Title |
Drosophila melanogaster larval male rep2 |
| Sample type |
RNA |
| |
|
| Source name |
larval male
|
| Organism |
Drosophila melanogaster |
| Characteristics |
strain: Oregon R
Sex: male
developmental stage: larval
tissue: whole organism
|
| Growth protocol |
Flies were grown at 22C on cornmeal media. Male and female larva were separately flash frozen on dry ice and stored at -80C. Virgin male and female newly eclosed flies were collected, aged for 3-5 days at 22C, and then frozen at -80C.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the samples using TRIzol (Invitrogen Inc, Carlsbad, CA), following the manufacter's instructions.
|
| Label |
Cy3
|
| Label protocol |
1000ng of total RNA in 10ul of RNase-free water was used to generate double-stranded cDNA using the Amino Allyl MessageAmp II aRNA Amplification kit (Ambion, Inc., Austin, TX), following the manufacter's instructions. The cDNA was then coupled to Cy3 reactive dye using the same kit, followed by a column purification using the kit-supplied equipment and reagents. Dye incorporation and cDNA yield were checked on a NanoDrop Spectrophotometer.
|
| |
|
| Hybridization protocol |
825ng of Cy3 labelled cDNA was fragmented and hybridized to the Agilent-018972 D. melanogaster Oligo Microarray 4x44K G2519F (GPL6385), following the manufacturer's instructions. After 17 hrs of hybridization at 65C, arrays were washed using the Agilent wash buffers per the manufacturer's instructions.
|
| Scan protocol |
Slides were scanned on the Axon 4000B scanner using the Agilent HD_GX_1Color setting (scan area 61x21.5 mm, scan resolution 5 um, and Green PMT set to XDR hi 100% and XDR low 10%).
|
| Description |
Biological replicate 2 of 3
|
| Data processing |
Scanned images were analyzed with the Agilent Feature Extraction Software version 10.5.1.1 (protocol GE1_105_DEC08) to obtain background substracted and spatially detrended processed signal intensities.
|
| |
|
| Submission date |
Aug 29, 2011 |
| Last update date |
Apr 08, 2012 |
| Contact name |
Richard Meisel |
| Organization name |
University of Houston
|
| Department |
Biology and Biochemistry
|
| Street address |
3455 Cullen Blvd
|
| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77204-5001 |
| Country |
USA |
| |
|
| Platform ID |
GPL6385 |
| Series (1) |
| GSE31722 |
Sex-biased gene expression in Drosophila melanogaster larvae |
|
