|
| Status |
Public on Feb 04, 2015 |
| Title |
EIHH001_REBOV_72hr_2_microRNA |
| Sample type |
RNA |
| |
|
| Source name |
IHH cells, REBOV treatment, 72 hours, replicate 2
|
| Organism |
Homo sapiens |
| Characteristics |
time: 72 hours
virus: REBOV
replicate: 2
|
| Treatment protocol |
Cells were infected with wild-type Ebola viruses at a multiplicity of infection of 0.5.
|
| Growth protocol |
IHH cells were maintained in DMEM medium with 5% FCS with L-glut, Anti-Anti, Na Pyruvate, and Non-essential AA
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were scraped into Trizol and total RNA was isolated from the Trizol extract.
|
| Label |
Cy3
|
| Label protocol |
Total RNA from each sample was labeled using miRNA Complete Labeling and Hyb Kit (v2.4, Agilent Technologies)
|
| |
|
| Hybridization protocol |
Sample labeling and array hybridization were performed according to the Agilent miRNA Complete Labeling and Hyb Kit protocol (Agilent Technology). Briefly, 500 ng of total RNA from each sample was labeled with Cy3-pCp. Labeled RNA was mixed with 4.5 μL 10 × Blocking Agent and 22.5 μL of 2X Hi-RPM Hybridization Buffer, heated at 100°C for 5 minutes, and then immediately transfered to an ice water bath for 5 minutes. All of the volume of the hybridization sample was dispensed into the gasket slide and assembled to the miRNA expression microarray slide. The slides were incubated for 55°C for 20 hours in an Agilent Hybridization Oven.
|
| Scan protocol |
The hybridized arrays were washed, fixed and scanned with Agilent DNA Microarray Scanner (part number G2505C).
|
| Data processing |
Scanned images were analyzed using Agilent Feature Extraction Software (v11.0.1.1). Quantile normalization and data processing performed using GeneSpring GX v12.0 software (Agilent Technologies).
|
| |
|
| Submission date |
Feb 03, 2015 |
| Last update date |
Aug 31, 2016 |
| Contact name |
Natalie Heller |
| E-mail(s) |
[email protected]
|
| Organization name |
PNNL
|
| Street address |
902 Battelle Blvd.
|
| City |
Richland |
| ZIP/Postal code |
99354 |
| Country |
USA |
| |
|
| Platform ID |
GPL19730 |
| Series (2) |
| GSE65573 |
Immortalized Human Hepatocyte (IHH) transcriptome response to wild-type Ebola viruses Zaire Ebola (ZEBOV '76) and Reston Ebola (REBOV '08) |
| GSE65575 |
Modeling Host Responses to Understand Severe Human Virus Infections |
|
