|
| Status |
Public on Dec 01, 2011 |
| Title |
0.5uM ecdy 4hr Glands rep2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Drosophila melanogaster, Oregon R, salivary glands, 0.5uM 20-hydroxy-ecdysone, 4hrs, Glands
|
| Organism |
Drosophila melanogaster |
| Characteristics |
cell line: Oregon R
genotype: db cn bw
tissue: salivary gland
developmental stage: late 3rd instar
agent: 0.5uM 20-hydroxy-ecdysone
time point: 4hrs
cell extraction: The sample had an [RNA] of 0.39 ug per uL and an OD ratio of 2.11. Details:0.5 mL Trizol100 uL chloroform300 mL isopropanol200 mL 75 percent EtOH-brought up in 10 uL Sigma 18 meg-ohm water
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cultures are grown to a minimum density of 5x10^6 cells per mL prior to extraction. Trizol is used at a ratio of 1 mL per 10^7 cells. Extraction procedure is as described by the manufacturer with centrifugation in a Sorvall SS-34 rotor. Final pellets are air dried and then resuspended in Sigma 18 meg-ohm water to obtain a final RNA concentation of no less than 4ug per uL.
|
| Label |
Cy5
|
| Label protocol |
CDMC Direct Labeling Protocol. See http://www.flyarrays.com/ under protocols.
|
| |
|
| Channel 2 |
| Source name |
Drosophila melanogaster, Oregon R, salivary glands, Ethanol control, 4hrs, Glands
|
| Organism |
Drosophila melanogaster |
| Characteristics |
cell line: Oregon R
genotype: db cn bw
tissue: salivary gland
developmental stage: late 3rd instar
agent: Ethanol control
cell extraction: The sample had an [RNA] of 0.35 ug per uL and an OD ratio of 2.13.Details: mL Trizol uL chloroform mL isopropanol mL 75 percent EtOH-brought up in 35uL Sigma 18 meg-ohm water
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cultures are grown to a minimum density of 5x10^6 cells per mL prior to extraction. Trizol is used at a ratio of 1 mL per 10^7 cells. Extraction procedure is as described by the manufacturer with centrifugation in a Sorvall SS-34 rotor. Final pellets are air dried and then resuspended in Sigma 18 meg-ohm water to obtain a final RNA concentation of no less than 4ug per uL.
|
| Label |
Cy3
|
| Label protocol |
CDMC Direct Labeling Protocol. See http://www.flyarrays.com/ under protocols.
|
| |
|
| |
| Hybridization protocol |
See http://www.flyarrays.com/ under protocols. Modifications by Scott:-use 5uL 50mM EDTA-precipitate probe for at least 1 hour-use 20uL DIG EasyHyb per humidifying well-final hyb volume is 80uL
|
| Scan protocol |
Images of the hybridized microarrays were obtained using a ScanArray 4000 scanner (Perkin-Elmer) and were quantified using QuantArray 3.0 software (Perkin-Elmer)
|
| Data processing |
Data were normalized using lowest sub-grid normalization (Yang, 2002) and Genetraffic Duo (Stratagene /Agilent) analysis software.
|
| |
|
| Submission date |
Sep 01, 2010 |
| Last update date |
Dec 01, 2011 |
| Contact name |
Zak Razak |
| E-mail(s) |
z.razak@utoronto.ca
|
| Phone |
905-569-4664
|
| Fax |
905-828-3792
|
| URL |
http://www.flyarrays.com
|
| Organization name |
Canadian Drosophila Microarray Centre
|
| Street address |
3359 Mississauga Rd
|
| City |
Mississauga |
| State/province |
Ontario |
| ZIP/Postal code |
L5L 1C6 |
| Country |
Canada |
| |
|
| Platform ID |
GPL1467 |
| Series (1) |
| GSE23928 |
Genome-wide examination of the transcriptional response to ecdysteroids 20-hydroxyecdysone and ponasterone A in Drosophila melanogaster |
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