|
| Status |
Public on Oct 31, 2017 |
| Title |
PHF20 gene was koncked down, biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
U87 cells infected with PHF20-shRNA lentivirus
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: U87 cell
cell type: human glioblastoma cell
transfection: shPHF20
|
| Treatment protocol |
U87 cells in 6-well plates were infected with lentiviral particles containing either shCON or shPHF20. The virus containing culture medium was replaced with fresh DMEM supplemented with 10% FBS at 12h after infection. 72h post lentivirus infection, two U87 cell groups were set up: KD (knockdown, cells infected with PHF20-shRNA lentivirus) and NC (negative control, cells infected with negative control-shRNA lentivirus).
|
| Growth protocol |
The cells were cultured in DMEM medium containing 10% FBS, 50U/mL penicillin and 50μg/mL streptomycin at 37℃ with 5% CO2 incubator. The cell line tested negative for any mycoplasma contamination.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA in the samples was isolated using Trizol Reagent according to the manufacture's protocol.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA were prepared according to the standard Affymetrix protocol(GeneChip® 3' IVT PLUS Reagent Kit, Affymetrix).
|
| |
|
| Hybridization protocol |
Total RNA was processed for double-strand cDNA synthesis, IVT and aRNA fragmentation with GeneChip 3’IVT Express Kit (Affymetrix, Santa Clara, CA, USA) according to the manufacturer’s instruction. Then, it was processed for hybridization at 45℃ for 17 h with The PrimeView™ Human Gene Expression Array (Affymetrix). The arrays were washed in the GeneChip Fluidic Station 450 (Affymetrix).
|
| Scan protocol |
The arrays were scanned by the GeneChip Scanner 3000 (Affymetrix).
|
| Data processing |
The raw data were normalized by the log scale robust multi-array analysis (RMA) method with the Expression Console software version 1.1 (Affymetrix).
|
| |
|
| Submission date |
Jan 17, 2017 |
| Last update date |
Oct 31, 2017 |
| Contact name |
Tianlong Liu |
| E-mail(s) |
[email protected]
|
| Organization name |
Xijing Hospital
|
| Department |
Pharmacy
|
| Street address |
No. 127 West Changle Road
|
| City |
Xi’an |
| ZIP/Postal code |
710032 |
| Country |
China |
| |
|
| Platform ID |
GPL16043 |
| Series (1) |
| GSE93680 |
Identification of genes and pathways that regulated by PHF20 via gene expression profile analysis in U87 cell with PHF20 silencing |
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