|
| Status |
Public on May 14, 2008 |
| Title |
GFP dsRNA-treated S2 cells (1) |
| Sample type |
RNA |
| |
|
| Source name |
Drosophila melanogaster Schneider2 (S2) cells , S2 cells treated with double-stranded RNA (dsRNA)
|
| Organism |
Drosophila melanogaster |
| Characteristics |
GFP dsRNA-treated S2 cells
|
| Treatment protocol |
S2 cells were soaked with dsRNA against GFP for 4 days.
After 4 days of initial soaking, cells were treated with a second round of dsRNA. Cells were harvested another 4 days later.
|
| Growth protocol |
S2 cells were grown at 25°C in Schneider's insect medium supplemented with 10% fetal bovine serum, 100U/ml penicillin and 100ug/ml streptomycin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with Trizol.
|
| Label |
-
|
| Label protocol |
2 ug of good quality total RNA were then labeled according to protocols recommended by manufacturers. Briefly, after reverse-transcription with an oligo-dT-T7 (Affymetrix), double stranded cDNA was generated with the supercript double stranded cDNA synthesis custom kit (Invitrogen Life Technologies, Carlsbad, CA). In an in vitro Transcription step (IVT) with T7 RNA polymerase (MessageAmp aRNA kit from Ambion, Austin, TX) the cDNA was linearly amplified and labeled with biotinylated nucleotides (Enzo Diagnostics, Farmingdale, NY).
|
| |
|
| Hybridization protocol |
10 ug of labeled and fragmented cRNA was then hybridized onto the Drosophila Genome 2.0 array (Affymetrix) for 16 hours at 45 degres celsius.
|
| Scan protocol |
Slides were stained and washed as indicated in the Antibody Amplification Stain for Eukaryotic Targets protocol using the Affymetrix Fluidics Station FS450. Arrays were then scanned with an Affymetrix Scanner 3000 and data was obtained using the Genechip® Operating Software 1.4
|
| Description |
-
|
| Data processing |
-
|
| |
|
| Submission date |
May 05, 2008 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Nicolas Robine |
| Organization name |
Sloan-Kettering Institute
|
| Department |
Developmental Biology
|
| Lab |
Dr. Eric Lai's Lab
|
| Street address |
430, East 67th Street, RRL 517
|
| City |
New York |
| State/province |
NY |
| ZIP/Postal code |
10065 |
| Country |
USA |
| |
|
| Platform ID |
GPL1322 |
| Series (1) |
| GSE11370 |
Two distinct mechanisms generate endogenous siRNAs from bidirectional transcription in Drosophila melanogaster. |
|
| Relations |
| Reanalyzed by |
GSE119084 |
