|
| Status |
Public on Nov 05, 2013 |
| Title |
ZHBTc4-EGFP vs Nr5a2-EGFP |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
ZHBTc4-EGFP clone #2-5
|
| Organism |
Mus musculus |
| Characteristics |
cell line: ZHBTc4 ES cell
transfection: pCAG-IP/EGFP
|
| Treatment protocol |
Doxycylin treated stable cells were harvest and extracted total RNA
|
| Growth protocol |
ES cells were cultured in the absence of feeder cells in Dulbecco's modified Eagles's medium(Gibco-BRL) supplied with 15% heat-inactivated fetal bovine serum(Sigma aldrich), 1mM sodium pyruvate(Sigma Aldrich), 10^-4M 2-mercaptoethanol, 1X nonessential amino acids(Sigma Aldrich), and 1000 IU/ml leukemia inhibitory factor (LIF; Millipore) on gelatin(0.2%) coated dishes.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
10 µg of total RNA were primed with 2 µl of 100 µM T16N2 DNA primer at 70°C for 10 min, then reversed transcribed at 42°C for 1 h in the presence of 400 U SuperScript II RTase (Invitrogen), and 100 µM each dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy3-label
|
| |
|
| Channel 2 |
| Source name |
ZHBTc4-Nr5a2-EGFP clone #26
|
| Organism |
Mus musculus |
| Characteristics |
cell line: ZHBTc4 ES cell
transfection: pCAG-IP/Nr5a2-EGFP
|
| Treatment protocol |
Doxycylin treated stable cells were harvest and extracted total RNA
|
| Growth protocol |
ES cells were cultured in the absence of feeder cells in Dulbecco's modified Eagles's medium(Gibco-BRL) supplied with 15% heat-inactivated fetal bovine serum(Sigma aldrich), 1mM sodium pyruvate(Sigma Aldrich), 10^-4M 2-mercaptoethanol, 1X nonessential amino acids(Sigma Aldrich), and 1000 IU/ml leukemia inhibitory factor (LIF; Millipore) on gelatin(0.2%) coated dishes.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions.
|
| Label |
Cy5
|
| Label protocol |
10 µg of total RNA were primed with 2 µl of 100 µM T16N2 DNA primer at 70°C for 10 min, then reversed transcribed at 42°C for 1 h in the presence of 400 U SuperScript II RTase (Invitrogen), and 100 µM each dATP, dTTP, dGTP, with 25 µM dCTP, 25 µM Cy3-label
|
| |
|
| |
| Hybridization protocol |
Oligoarray control targets and hybridization buffer (Agilent In Situ Hybridization Kit Plus) were added, and samples were applied to microarrays enclosed in Agilent SureHyb-enabled hybridization chambers. After hybridization, slides were washed sequential
|
| Scan protocol |
Scanned on an Agilent G2565AA scanner.
|
| Description |
Stable EGFP or Nr5a2-EGFP gene transfected ZHBTc4 ES cells, Monoclonal cell line, Doxycyclin treatment before harvest
|
| Data processing |
Agilent Feature Extraction Software (v 9.3.2.1) was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
Nov 04, 2013 |
| Last update date |
Nov 05, 2013 |
| Contact name |
kyeng won choi |
| E-mail(s) |
[email protected]
|
| Organization name |
Sogang
|
| Department |
Lifescience
|
| Lab |
MCBio
|
| Street address |
Sinsu
|
| City |
Mapo |
| State/province |
Seoul |
| ZIP/Postal code |
121742 |
| Country |
South Korea |
| |
|
| Platform ID |
GPL11202 |
| Series (1) |
| GSE52034 |
ZHBTc4 ES cells stably expressing EGFP vs Nr5a2-EGFP or Oct4-EGFP vs Nr5a2-EGFP |
|
