|
| Status |
Public on Nov 01, 2007 |
| Title |
H16N2 cell treated with EZH2 siRNA Replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Total RNA from EZH2 siRNA-treated H16N2 cells labeled with Cyanine-5 (red).
|
| Organism |
Homo sapiens |
| Characteristics |
H16N2
EZH2 siRNA-transfected
Age: day 2
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
30µg of total RNA were primed with 6 µl of poly-dT primers for 10min, then reverse-transcribed at 42°C for 1 h in the presence of 400 U SuperScript II RTase (Invitrogen), and 100 µM each dATP, dCTP, dGTP, with 25 µM dTTP, 25 µM aadUTP; the reaction mix were then cleaned and concentrated by Microcon filter, and labelled with Cy3 or Cy5 dyes at RT for 1hour
|
| |
|
| Channel 2 |
| Source name |
Total RNA from luciferase siRNA-treated H16N2 cells labeled with Cyanine-3 (green).
|
| Organism |
Homo sapiens |
| Characteristics |
luciferase siRNA-transfected
Age: day 2
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
30µg of total RNA were primed with 6 µl of poly-dT primers for 10min, then reverse-transcribed at 42°C for 1 h in the presence of 400 U SuperScript II RTase (Invitrogen), and 100 µM each dATP, dCTP, dGTP, with 25 µM dTTP, 25 µM aadUTP; the reaction mix were then cleaned and concentrated by Microcon filter, and labelled with Cy3 or Cy5 dyes at RT for 1hour
|
| |
|
| |
| Hybridization protocol |
human Cot DNA and yeast tRNA were added to labelled cDNA and hybridized in SSC/SDS buffer to arrays
|
| Scan protocol |
Scanned on an Affymetrix Axon scanner
Images were quantified using Genepix6.0
|
| Description |
Biological replicate 2 of 3. EZH2-siRNA-treated vs. luciferase siRNA-treated control
|
| Data processing |
median normalized, background subtracted data obtained
|
| |
|
| Submission date |
Jun 15, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Jindan Yu |
| E-mail(s) |
jindan-yu@northwestern.edu
|
| Organization name |
Northwestern University
|
| Department |
Medicine - Hem/Onc
|
| Lab |
Yu
|
| Street address |
303 E. Superior St.
|
| City |
Chicago |
| State/province |
IL |
| ZIP/Postal code |
60611 |
| Country |
USA |
| |
|
| Platform ID |
GPL2013 |
| Series (1) |
| GSE8145 |
human prostate and breast epithelial cell lines with EZH2 siRNA interference |
|
