|
| Status |
Public on May 21, 2013 |
| Title |
NetFra-replicate-2 |
| Sample type |
RNA |
| |
|
| Source name |
Drosophila wing discs ectopically expressing NetA+Fra
|
| Organism |
Drosophila melanogaster |
| Characteristics |
tissue: wing imaginal disc
age: 3rd instar
genotype: 70FLP/+; GAL4-Act5C(FRT.CD2).P}S UAS-GFP/UAS-NetA UAS-fra
|
| Treatment protocol |
Wing discs were dissected out of 3rd instar larvae in PBS, snap frozen in liguid nitrogen, and stored at -80°C for subsequent RNA extraction.
|
| Growth protocol |
Eggs were laid during a four hour time window. Animals were reared at room temperature on standard fruit fly culturing media. Larvae were heat shocked at 37°C for 20 minutes 48 hours after egg laying to induce clone formation. Animals were then allowed to grow at room temperature until emerging as 3rd instar larvae.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA were extracted using Trizol (Invitrogen) per the manufacturer's instructions. RNA sample integrity was verified with a 2100 Bioanalyzer (Agilent Technologies, Inc.).
|
| Label |
biotin
|
| Label protocol |
Target aRNA were generated and prepared using the GeneChip 3'IVT Express kit (Affymetrix, Inc.) according to the manufacturer's instructions. Briefly, reverse transcriptase and an oligo(dT) primer were used to synthesize first-strand cDNA. Second-strand cDNA was synthesized using DNA polymerase. Biotin-modified amplified RNA (aRNA) was generated through in vitro transcription. The aRNA was purified, fragmented, then hybridized in a GeneChip Hybridization Oven 640 at 45 degrees C for 16 hours.
|
| |
|
| Hybridization protocol |
aRNA were hybridized to the Affymetrix GeneChip Drosophila Genome 2.0 in a GeneChip Hybridization Oven 640 according to the manufacturer's instructions. The GeneChips were washed and stained in an Affymetrix Fluidics Station 450.
|
| Scan protocol |
GeneChips were scanned with a GeneChip 3000 7G Scanner.
|
| Description |
Gene expression data for Drosophila third instar wing imaginal discs ectopically expressing NetA+Fra
|
| Data processing |
MAS 5.0 signal
|
| |
|
| Submission date |
May 20, 2013 |
| Last update date |
May 21, 2013 |
| Contact name |
Molly Duman Scheel |
| Organization name |
Indiana University School of Medicine
|
| Street address |
1234 Notre Dame Ave.
|
| City |
South Bend |
| State/province |
IN |
| ZIP/Postal code |
46617 |
| Country |
USA |
| |
|
| Platform ID |
GPL1322 |
| Series (1) |
| GSE47113 |
Gene expression profile of Drosophila third instar larval wing imaginal discs overexpressing netrinA (netA) and frazzled (fra). |
|
