|
| Status |
Public on Aug 26, 2011 |
| Title |
325T_Macro |
| Sample type |
RNA |
| |
|
| Source name |
Macro Samples
|
| Organism |
Homo sapiens |
| Characteristics |
disease state: Tumor
cell type: Adeno carcinoma
|
| Extracted molecule |
total RNA |
| Extraction protocol |
50mg to 80mg of snap frozen lung tissue was added to a tube containing 1 ml of ice cold trizol and homogenized until tissue was totally homogenized. After incubation for 5 min at room temperature, 0.2ml of chloroform was added; tube was shaked vigorously by hand for 15 sec and then incubated for 3 min at room temperature. Then samples were centrifuged for 20 min at 12000g at 4C. Aqueous phase was transfered to a clean 1.5-ml tube and 1 x volume of 70% ethanol was added into the aqueous phase. Sample was applied to the Rneasy column and centrifuged at 8000g for 30 sec. Further procedure was done using Rneasy mini kit according to the manufactures suggestions including an optional 30 min DNase I treatment.
|
| Label |
biotin
|
| Label protocol |
100ng total RNA was amplified using Ambion WT Expression Kit following the manufacturer’s instructions. 5.5 µg of amplification products were labeled with Affymetrix WT Terminal Labeling Kit hybridized to Affymetrix HG Gene 1.0ST chips using Affymetrix Hybridization Control Kit and washed and stained using Affymetrix Wash and Stain Kit. All steps were done following the manufacturer’s instructions.
|
| |
|
| Hybridization protocol |
Affymetrix protocol
|
| Scan protocol |
Affymetrix protocol
|
| Data processing |
Affymetrix Expression Console software and RMA (Robust Multichip Analysis) normalization was performed to generate the CHP files.
|
| |
|
| Submission date |
Aug 19, 2011 |
| Last update date |
Apr 26, 2012 |
| Contact name |
Miao Shi |
| E-mail(s) |
[email protected]
|
| Phone |
1-718-678-1035
|
| Organization name |
Albert einstein College of Medicine
|
| Street address |
1301 Morris Park ave
|
| City |
Bronx |
| State/province |
NY |
| ZIP/Postal code |
10461 |
| Country |
USA |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE31552 |
Expression Data from human Lung tissue of Patients with Non Small Cell Lung Cancer (NSCLC) |
|
