|
| Status |
Public on Aug 12, 2010 |
| Title |
Salivary gland vs Brain and Disc |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
salivary glands
|
| Organism |
Drosophila melanogaster |
| Characteristics |
extraction method: hand dissected
tissue: salivary gland
developmental stage: 3rd instar larvae collected 70-75 hours after egg laying
genome/variation: wild-type
strain: yw67c23
cell cycle status: Endocycling tissue
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Drosophila adults yw67C23 strain were put on food for 6 hours, and the resulting population of 70-75 hours old larvae (feeding-stage 3rd instar) were used for hand dissection of salivary gland tissues.
|
| Label |
Cy3
|
| Label protocol |
Total RNA was reverse transcribed and fluorescently labeled with Cy3 (Amersham) using Amino Allyl MessageAmp™ II aRNA Amplification Kit (Ambion, Austin, TX 78744), according to the manufacturer’s instructions.
|
| |
|
| Channel 2 |
| Source name |
brain-disc
|
| Organism |
Drosophila melanogaster |
| Characteristics |
extraction method: hand dissected
tissue: brain-disc complexes
developmental stage: 3rd instar larvae collected 70-75 hours after egg laying
genome/variation: wild-type
strain: yw67c23
cell cycle status: Mitotic cycling tissue
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Drosophila adults yw67C23 strain were put on food for 6 hours, and the resulting population of 70-75 hours old larvae (feeding-stage 3rd instar) were used for hand dissection of brain-disc complexes.
|
| Label |
Cy5
|
| Label protocol |
Total RNA was reverse transcribed and fluorescently labeled with Cy5 (Amersham) using Amino Allyl MessageAmp™ II aRNA Amplification Kit (Ambion, Austin, TX 78744), according to the manufacturer’s instructions.
|
| |
|
| |
| Hybridization protocol |
Methods were as described (Bogart et al., 2006) and details can be found at (https://dgrc.cgb.indiana.edu/microarrays/support/protocols.html).
|
| Scan protocol |
Hybridized arrays were scanned by GenePix 4100A (Axon Instruments, Union City, CA).
|
| Description |
This is the average value for three biological replicate experiments. Individual values for each independent experiment is provided in the supplementary file.
|
| Data processing |
The data was extracted with Axon GenePix Pro 5 image analysis software. The spot sizes and intensities quantified by the software and automatically flagged spot qualities were followed manual examination. Abnormal shape spots or spots with high local background or spots that were quantified due to false intensity caused by dust were flagged bad and discarded. The normalization of the intensity values from the two channels was performed using global normalizations. After normalizations data points with background subtracted median intensity signals <60% than two standard deviations (SD) above the overall background intensity in both channels were discarded. Further data analyses were performed using data analysis software Acuity 4 (Axon) and Microsoft Excel. Data points were averaged from replicate spots and inconsistent data points were removed manually.
|
| |
|
| Submission date |
Nov 03, 2009 |
| Last update date |
Aug 11, 2010 |
| Contact name |
Brian R. Calvi |
| E-mail(s) |
bcalvi@indiana.edu
|
| Phone |
812-855-5450
|
| Fax |
812-855-6705
|
| Organization name |
Indiana University
|
| Department |
Biology
|
| Street address |
1001 E. 3rd St.
|
| City |
Bloomington |
| State/province |
IN |
| ZIP/Postal code |
47405 |
| Country |
USA |
| |
|
| Platform ID |
GPL9528 |
| Series (1) |
| GSE19029 |
Gene expression profiling of endocycling cells vs mitotic cells |
|
