|
| Status |
Public on Dec 31, 2016 |
| Title |
HRSV at 0H, biological rep2 |
| Sample type |
RNA |
| |
|
| Source name |
Mock infected Hep2
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Hep2 cells
|
| Treatment protocol |
The cells were infected with HRSV MOI=3 in DMEM + Glutamax (Gibco) with 2% Fetal Bovine Serum (FBS) and 100 units/ml of Penicillin/Sreptomycin (Gibco), at 37˚C, 5% CO2
|
| Growth protocol |
The cells were grown in DMEM L929 (30%) cell condition media (Gibco) and 100 units/ml of Penicillin/Sreptomycin (Gibco), at 33˚C, 5% CO2
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA extraction was performed using RNAeasy MiniKit (Qiagen) according to the manufacturer's instruction at 4˚C
|
| Label |
biotin
|
| Label protocol |
Biotinylated mRNA were prepared according to the standard Affymetrix protocol from 150 ng total RNA (Expression Analysis Technical Manual, 2009, Affymetrix).
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 ug of aRNA were hybridized for 16 hr at 45˚C on Human U133 2.0 Genome Array. GeneChips were washed and stained in the Affymetrix Fluidics Station 400.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000
|
| Description |
Gene expression data from mock infected Hep2 rep 2
Hep2_HRSV_0H-2
|
| Data processing |
The data were analyzed with GeneChip Operating Software 5.0 (GCOS 5.0) using Affymetrix default analysis settings. Further analysis was done in GeneSpring 11.0. The normalization was done using internal and external control genes as assigned by Affymetrix. The target intensity of each array on a given time point was first filtered by the flagging of Present (P) in 3 replicates at one time-point, which then further calculated in fold change up- or downregulation with respect to the signal intensity in MOCK infected cells, after analyzed by ANOVA with p value ≤ 0.05.
|
| |
|
| Submission date |
Nov 04, 2011 |
| Last update date |
Dec 31, 2016 |
| Contact name |
Richard J Sugrue |
| E-mail(s) |
RJSUGRUE@ntu.edu.sg
|
| Organization name |
Nanyang Technological University
|
| Department |
Divison of Molecular and Cell Biology
|
| Street address |
60 Nanyang Drive
|
| City |
Singapore |
| ZIP/Postal code |
637551 |
| Country |
Singapore |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE33485 |
Host cell gene expression profile in Human respiratory syncytial virus (HRSV) infected Hep2 cells at 4, 8, 12 and 15 hours post infection |
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