|
| Status |
Public on Feb 13, 2017 |
| Title |
Day 0 replicate 1 |
| Sample type |
RNA |
| |
|
| Source name |
Archived plasma_day 0_post-exposure to aerosolized Ebola virus
|
| Organism |
Macaca mulatta |
| Characteristics |
tissue: plasma
day post-exposure: 0
|
| Treatment protocol |
Plasma was mixed with 3 volumes of TRIzol LS after separation from whole blood.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
After TRIzol phase separation, the aqueous phase was used as input to the miRNeasy Mini Kit for total RNA extraction. 5 μg of molecular biology-grade glycogen was added as an RNA co-precipitant.
|
| Label |
SYBR Green
|
| Label protocol |
RT-PCR assays were performed using the Exiqon microRNA Ready-to-Use PCR Human panel I+II V3.R (catalog number 203612). cDNA synthesis was performed using the cDNA Synthesis Kit II using 8 μL RNA for each 40 μL sample reaction.
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
Control
Day0_1
|
| Data processing |
Cq values were calculated using the second derivative method of the integrated Roche LightCycler 480 software version 1.5.1.
Cq values were imported into GenEx 6 Academic Enterprise software for further analysis.
MiRNA expression levels were calculated using the ΔΔCq method with global normalization. Briefly, interplate and spike-in calibrations were performed, followed by removal of samples with Cq > 39.4 and miRNAs that were not detected in 5/6 replicates. Global normalization of Cq values was performed by using the mean of all samples with Cq < 36 (per day). Relative quantities of each miRNA were calculated with respect to the average expression level on day 0, then data were log2 transformed and compared using a paired t-test.
Matrix normalized sheet reports values after interplate calibration and global normalization.
Fold change sheet reports log2 transformed test/control (day 7/day 0) values. Non-normalized values, Normalized values and and fold-change data are available on the series record.
|
| |
|
| Submission date |
Dec 30, 2014 |
| Last update date |
Feb 13, 2017 |
| Contact name |
Janice Duy |
| Organization name |
U.S. Army Medical Research Institute of Infectious Diseases
|
| Department |
Diagnostics Systems Division
|
| Street address |
1425 Porter St.
|
| City |
Frederick |
| State/province |
MD |
| ZIP/Postal code |
21702 |
| Country |
USA |
| |
|
| Platform ID |
GPL17107 |
| Series (1) |
| GSE64579 |
MicroRNA expression profiling of one rhesus macaque at day 0 and day 7 post-exposure to aerosolized Ebola virus |
|
