|
| Status |
Public on Apr 01, 2008 |
| Title |
ST11-196, TERT and SV40EA transfected, Agilent |
| Sample type |
RNA |
| |
|
| Source name |
human fetal lung fibroblast TIG-1 clone ST11 transfected with TERT and SV40EA
|
| Organism |
Homo sapiens |
| Characteristics |
human fetal lung fibroblast TIG-1 clone ST11 at 196 PDL transfected with TERT and SV40EA
|
| Biomaterial provider |
Health Science Research Resources Bank, Tokyo, Japan
|
| Treatment protocol |
TERT expression vector and SV40 early antigens (SV40EA) expression vector were co-transfected to TIG-1 and isolated as a G418 resistant clone. Exponentially growing cultured cells were collected using trypsin followed by rinse with PBS, and freezed until use.
|
| Growth protocol |
Cells were cultured in Dulbecco's modified Eagle's minimal essential medium (DMEM; Sigma-Aldrich Japan, Tokyo, Japan) containing 10% heat-inactivated fetal bovine serum (FBS; BioWhittaker, Verviers, Belgium) in a humidified atmosphere of 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the frozen cultured cell pellets using QIAGEN RNeasy mini kit (QIAGEN, Inc., Valencia, CA), respectively, according to the manufacturer's protocols. The quality of the RNA was checked using Agilent Technologies 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
The cDNA was generated from 0.6 microgram of total RNA using reverse transcriptase and a T7 primer using Agilent Low RNA Input Linear Amplification Kit PLUS, One-color (Agilent). cRNA (complementary RNA) was generated via an in vitro transcription reaction using T7 RNA polymerase and cyanine 3-labeled CTP
|
| |
|
| Hybridization protocol |
The synthesized 1.5 microgram of cRNA, quantified by spectrometry and qualified using an Agilent 2100 Bioanalyzer, was then fragmented and hybridized to each array. After 17hr hybridization, arrays were washed according to the manufacturer's recommended protocol.
|
| Scan protocol |
Each microarray was scanned using an Agilent DNA Microarray Scanner.
|
| Description |
immortal lifespan, transformed
|
| Data processing |
Expression levels were quantified by Agilent Feature Extraction software ver. 9.1. and normalized to the median expression value of the whole array spots using GeneSpringTM GX (Agilent Technologies).
|
| |
|
| Submission date |
Sep 18, 2007 |
| Last update date |
Aug 14, 2011 |
| Contact name |
Keiko Hiyama |
| E-mail(s) |
khiyama@hiroshima-u.ac.jp
|
| Phone |
81-82-257-5841
|
| Fax |
81-82-256-7105
|
| Organization name |
Hiroshima University
|
| Department |
Research Institute for Radiation Biology and Medicine
|
| Lab |
Dept. Translational Cancer Research
|
| Street address |
1-2-3 Kasumi, Minami-ku
|
| City |
Hiroshima |
| State/province |
Hiroshima |
| ZIP/Postal code |
734-8553 |
| Country |
Japan |
| |
|
| Platform ID |
GPL4133 |
| Series (1) |
| GSE9077 |
Expression profiles of immortal lung fibroblasts |
|
