|
| Status |
Public on Jul 20, 2013 |
| Title |
E2_H008 |
| Sample type |
RNA |
| |
|
| Source name |
normal bronchial epithelial cell
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Beas-2B
genotype/variation: pEGFPN1-ECPsp
|
| Extracted molecule |
total RNA |
| Extraction protocol |
On average, 50 7 um-thick cryostat sections were prepared for the extraction of total RNA from tumor cell-rich areas, which were identified by a surgical pathologist (YY) using every 10th section stained by May-Giemsa. RNA was extracted using Trizol.
|
| Label |
[33P]dCTP
|
| Label protocol |
Five mg of total RNA was reverse-transcribed using oligo-dT primers (Invitrogen) and SuperScript II reverse transcriptase (Invitrogen) in the presence of 100 mCi of [33P]dCTP (Amersham Bioscience, Piscataway, NJ) according to the instructions for the Gene
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| |
|
| Hybridization protocol |
The GeneFilters were prehybridized for 2 h at 51C with 0.5 mg/ml of poly-dA (Invitrogen) and 0.5 mg/ml Cot-1 DNA (Invitrogen) in 10 ml of AlkPhos DIRECT hybridization buffer (Amersham Bioscience) and then hybridized for 17 h at 51C with the denatured radi
|
| Scan protocol |
The arrays were then exposed for 2 hours to an Imaging Plate and scanned at 25-mm resolution with a BAS5000 phosphoimager (Fuji Photo Film), images of the hybridized arrays were processed with L Process (Fuji Photo Film) and signal intensities quantified
|
| Data processing |
The raw data were re-scaled to account for the differences in individual hybridization intensities. We employed a rank-invariant scaling method to select genes (Tseng et al., 2001), which were then used for fitting of a non-linear normalization curve. Rosetta Resolver System (Rosetta Biosoftware) was used to normalize the data.
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| |
|
| Submission date |
Jul 05, 2012 |
| Last update date |
Jul 20, 2013 |
| Contact name |
Hao-Teng Chang |
| E-mail(s) |
htchang@mail.cmu.edu.tw
|
| Organization name |
China Medical University
|
| Street address |
No.91, Hsueh-shih Rd. N. Dist.
|
| City |
Taichung Ciy |
| ZIP/Postal code |
40402 |
| Country |
Taiwan |
| |
|
| Platform ID |
GPL15493 |
| Series (1) |
| GSE39122 |
Gene expression-based, inflammatory response prediction by bronchial epithelial cell line treated with signal peptide of eosinophil cationic protein |
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