|
| Status |
Public on Aug 09, 2007 |
| Title |
transfected HeLa cells; mock versus siGFP 2a |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source Name |
Total RNA from mock transfected HeLa cells labeled with Cyanine-3
|
| Organism(s) |
Homo sapiens |
| Characteristics |
HeLa cell line (CCL-2)
|
| Treatment protocol |
Cells were transfected using the transfection reagent Effentene according to the manufacturer's protocoll.
|
| Growth protocol |
Cells were grown in an incubator with 95% relative air humidity and 5% CO2 in the adequate medium supplemented with 10% FCS, L-glutamine and antibiotics.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
Samples were labeled with fluorochrome by the Klenow fragment using the Bioprime Kit (Invitrogen, Karlsruhe, Germany).
|
| |
|
| Channel 2 |
| Source Name |
Total RNA from siGFP transfected HeLa cells labeled with Cyanine-5
|
| Organism(s) |
Homo sapiens |
| Characteristics |
HeLa cell line (CCL-2)
|
| Treatment protocol |
Cells were transfected using the transfection reagent Effentene according to the manufacturer's protocoll.
|
| Growth protocol |
Cells were grown in an incubator with 95% relative air humidity and 5% CO2 in the adequate medium supplemented with 10% FCS, L-glutamine and antibiotics.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
Samples were labeled with fluorochrome by the Klenow fragment using the Bioprime Kit (Invitrogen, Karlsruhe, Germany).
|
| |
|
| |
| Hybridization protocol |
Purified, dye-labeled cDAN was mixed with Ultra-Hyb buffer (Ambion), agitated for 60min at 60°, then for 10min at 70°C on a thermo mixer and subsequently applied to pre-heated microarrays mounted in a GeneTAC Hybridisation Station. After hybridization slides were washed sequential.
|
| Scan protocol |
Microarrays were scanned at 5 mm resolution and variable PMT voltage to abtain maximal signal intensities with <0.1% probe saturation, a count ratio of 0.8-1.2 (Cy5/Cy3) and maximal congruence of histogram curves, using a GenePix 4000B microarray scanner.
|
| Description |
Colour-switch 2a
|
| Data processing |
Dta summaries on the spot intensities are provided by GenePix Pro, Version 5.1, using local background correction. This includes the mean and the median pixel intensities at each scan wavelength for both feature and background pixels.
|
| |
|
| Submission date |
Aug 03, 2007 |
| Contact name |
Cordula Tschuch |
| E-mail(s) |
c.tschuch@dkfz.de
|
| Phone |
+49-6221-424594
|
| Organization name |
DKFZ
|
| Street address |
Neuenheimer Feld 580
|
| City |
Heidelberg |
| ZIP/Postal code |
69120 |
| Country |
Germany |
| |
|
| Platform ID |
GPL5704 |
| Series (1) |
| GSE8680 |
HEK and HeLa cells: transfection of siRNA directed against GFP versus mock transfection |
|
