|
| Status |
Public on Mar 31, 2009 |
| Title |
Synchronized HCT116_Nocodazole_8h |
| Sample type |
RNA |
| |
|
| Source name |
Synchronized HCT116, Nocodazole, 8h
|
| Organism |
Homo sapiens |
| Characteristics |
Cell line: HCT116
Treatment: Nocodazole
Time course: 8h
|
| Treatment protocol |
Synchronous culture was obtained by incubating cells for 19 h in 2 mM of thymidine, followed by a 9-h incubation in normal medium and a second 16-h incubation in thymidine (2 mM). Cells were washed with normal medium followed by treatment with DMSO for 0, 2, 4, 6, 7, 8, 9, and 10 h as a control or 0.1 mg/ml nocodazole (Sigma-Aldrich) for 7, 8, 9, and 10 h.
|
| Growth protocol |
The HCT116 colorectal cancer cell line (ATCC) was grown in McCoy's 5A medium modified (Sigma-Aldrich) with 10% FBS (JBS) and maintained at 37°C and 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated with RNeasy kits (Qiagen) according to the manufacturers' instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the manufacturer's instructions (Affymetrix Expression Analysis Technical Manual, Rev. 4).
|
| |
|
| Hybridization protocol |
The cRNA products were fragmented, heated at 99 degrees C for 5 min and hybridized for 16 h at 45 degrees C. The microarrays were then washed at low (6X SSPE) and high (100mM MES, 0.1M NaCl) stringency and stained with streptavidin-phycoerythrin. Fluorescence was amplified by adding biotinylated anti-streptavidin and an additional aliquot of streptavidin-phycoerythrin stain.
|
| Scan protocol |
Scanning was carried out using the GeneChip Scanner and image analysis was performed using GeneChip Operating Software.
|
| Description |
N/A
|
| Data processing |
GCRMA
|
| |
|
| Submission date |
Dec 22, 2008 |
| Last update date |
Mar 31, 2009 |
| Contact name |
Hideaki Mizuno |
| E-mail(s) |
mizunohda@chugai-pharm.co.jp
|
| URL |
http://researchmap.jp/mizuno
|
| Organization name |
Chugai Pharmaceutical Co., Ltd.
|
| Department |
Discovery research dept.
|
| Lab |
Kamakura research labs.
|
| Street address |
200 Kajiwara
|
| City |
Kamakura |
| State/province |
Kanagawa |
| ZIP/Postal code |
247-8530 |
| Country |
Japan |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE14103 |
Synchronized HTC116 cells: time course |
|
