|
| Status |
Public on Nov 19, 2014 |
| Title |
THC only 60D [HE44-60D] |
| Sample type |
RNA |
| |
|
| Source name |
THC only 60D
|
| Organism |
Macaca mulatta |
| Characteristics |
tissue: Intact Duodenum
days post siv infection (dpi): 60 D
|
| Treatment protocol |
8 animals were infected intravenously with 100 TCID50 SIVmac251 and 4 (3 THC only and 1 vehicle only) remained uninfected
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA (small and large) was extracted using the miRNeasy Mini kit (Qiagen). On-column DNase digestion was also performed to eliminate contaminating genomic DNA.
|
| Label |
n/a
|
| Label protocol |
microRNA profiling was performed using the TaqMan® OpenArray® Human microRNA panels (Life Tech) following the manufacturer’s instructions. Reverse transcription was first performed from 350 ng of total RNA using the microRNA reverse transcription kit. 2.5 µL of cDNA was preamplified using the TaqMan® PreAmp Master Mix kit. Preamplified products were first diluted and then loaded onto to TLDA plates. TLDA plates were run on the ABI 7900 HT Fast PCR system following the manufacturer’s recommended thermal cycling conditions.
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
Test
|
| Data processing |
QuantStudio run files representing all treatment groups and timepoints were analyzed using ExpressionSuite Software (Life Tech)
miRNA expression data was analyzed by normalizing to three endogenous controls (RNU44, RNU48 and snoU6)
In all experiments, the CT upper limit was set to 28 meaning that all miRNA detectors with a CT value greater than or equal to 28 were excluded.
Due to the small sample size and exploratory nature of the study multiple comparisons correction was not applied.
ExpressionSuite software automatically calculates all deltadeltaCT based fold-change calculations and provides p values from the uploaded QuantStudio files.
Fold Change worksheet reports test/control (i.e., THC or VEH/SIV, THC/SIV vs uninfected control) ratios.
|
| |
|
| Submission date |
Sep 22, 2014 |
| Last update date |
Nov 19, 2014 |
| Contact name |
Mahesh Mohan |
| E-mail(s) |
mmohan@txbiomed.org
|
| Organization name |
Southwest National Primate Research Center
|
| Department |
Host Pathogen Interaction Program
|
| Lab |
12/106
|
| Street address |
8715 West Military Road
|
| City |
San Antonio |
| State/province |
Texas |
| ZIP/Postal code |
78227 |
| Country |
USA |
| |
|
| Platform ID |
GPL17837 |
| Series (1) |
| GSE61654 |
miRNA profiling of Macaca mulatta intact duodenum samples following chronic Delta9 Tetrahydrocannabinol (Δ9-THC) treatment to SIV infected rhesus macaques |
|
