|
| Status |
Public on Feb 09, 2012 |
| Title |
G1/S-synchronized CL1-5 cells |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
CL1-5/AS2neo G1/S
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: CL1-5/AS2neo (control)
|
| Treatment protocol |
For double thymidine block and release experiments, cells were treated with 2 mM thymidine (Sigma) for 19 h and released for 9 h, followed by the second 16 h treatment before being harvested.
|
| Growth protocol |
CL1-5/AS2neo (control) and CL1-5/AS2neo-Slug stable cells were cultured in RPMI medium containing 10% FBS, l-glutamine, and 400ug/ml G418
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy3
|
| Label protocol |
0.2 μ g of total RNA was amplified by a Low Input Quick-Amp Labeling kit (Agilent Technologies, USA) and labeled with Cy3 or Cy5 (CyDye, Agilent Technologies, USA) during the in vitro transcription process.
|
| |
|
| Channel 2 |
| Source name |
CL1-5/AS2neo-Slug G1/S
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: CL1-5/AS2neo-Slug-WT stable cells
|
| Treatment protocol |
For double thymidine block and release experiments, cells were treated with 2 mM thymidine (Sigma) for 19 h and released for 9 h, followed by the second 16 h treatment before being harvested.
|
| Growth protocol |
CL1-5/AS2neo (control) and CL1-5/AS2neo-Slug stable cells were cultured in RPMI medium containing 10% FBS, l-glutamine, and 400ug/ml G418
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA extracted using Trizol following manufacturer's instructions
|
| Label |
Cy5
|
| Label protocol |
0.2 μ g of total RNA was amplified by a Low Input Quick-Amp Labeling kit (Agilent Technologies, USA) and labeled with Cy3 or Cy5 (CyDye, Agilent Technologies, USA) during the in vitro transcription process.
|
| |
|
| |
| Hybridization protocol |
0.3 μ g of Cy-labled cRNA was fragmented to an average size of about 50-100 nucleotides by incubation with fragmentation buffer at 60°C for 30 minutes. Correspondingly fragmented labeled cRNA is then pooled and hybridized to a Agilent SurePrint G3 Human GE 8×60K Microarray (Agilent Technologies, USA) at 65°C for 17 h.
|
| Scan protocol |
Scanned on an Agilent microarray scanner (Agilent Technologies, USA)
|
| Description |
cells were synchronized with double thymidine at G1/S phase
|
| Data processing |
Scanned images are analyzed by Feature extraction 10.5.1.1 software (Agilent Technologies, USA), an image analysis and normalization software is used to quantify signal and background intensity for each feature, substantially normalized the data by rank-consistency-filtering LOWESS method.
|
| |
|
| Submission date |
Feb 08, 2012 |
| Last update date |
Feb 10, 2012 |
| Contact name |
Tse-Ming Hong |
| E-mail(s) |
htmjimmy@gmail.com
|
| Organization name |
Academia Sinica
|
| Street address |
128 Academia Road, Section 2, Nankang
|
| City |
Taipei |
| ZIP/Postal code |
115 |
| Country |
Taiwan |
| |
|
| Platform ID |
GPL13607 |
| Series (1) |
| GSE35617 |
The invasion promoter Slug is a critical cell cycle regulator |
|
