|
| Status |
Public on Mar 15, 2012 |
| Title |
A375 siRNA-PCNA 48h |
| Sample type |
RNA |
| |
|
| Source name |
A375 cells that were transfected by siRNA targeting the corresponding gene
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: A375
knockdown: siRNA-PCNA
|
| Treatment protocol |
A375 cells were seeded in 6-well plates. 20h after seeding when the cells reached 30% confluence, they were transfected with 100nm of siRNA with 30μl Oligofectamine accroding to the manufacture's instruction. After 48h, the cells were harvested and RNAs were used for the microarray analysis.
|
| Growth protocol |
A375 cells were grown in DMEM medium containing 10% FBS at 37 ˚C in a fully humidified atmosphere containing 5% CO2
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Trizol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol from 4 ug total RNA (Expression Analysis Technical Manual, 2001, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of labelled cRNA was hybridized for 16 hr at 45°C @ 60 RPM on Affymetrix GeneChip Human Genome U133 Plus 2.0 Arrays. GeneChips were washed and stained in the Affymetrix Fluidics Station 450
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 7G
|
| Description |
Gene expression profile of A375 cells 48 h after the corresponding gene was knockdown by siRNA was analyzed using Affymetrix Human Genome U133 plus 2.0 microarrays
|
| Data processing |
The data were analyzed with R using Affymetrix default analysis settings and RMA as normalization method.
|
| |
|
| Submission date |
Aug 19, 2011 |
| Last update date |
Mar 15, 2012 |
| Contact name |
Li Wang |
| E-mail(s) |
[email protected]
|
| Organization name |
University of Auckland
|
| Department |
Molecular Medicine & Pathology
|
| Lab |
Print Laboratory
|
| Street address |
85 Park Rd
|
| City |
Auckland |
| ZIP/Postal code |
1023 |
| Country |
New Zealand |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE31534 |
Gene expression profile in A375 melanoma cells after 45 functionally important molecules were knocked down using siRNA |
|
