|
| Status |
Public on Nov 08, 2023 |
| Title |
HEV-3c infected (SAMPLE 5) |
| Sample type |
RNA |
| |
|
| Source name |
Liver
|
| Organism |
Sus scrofa |
| Characteristics |
tissue: Liver tissue
treatment: HEV-3c
time: 8 days post-infection
moi: 10^6 copies HEV RNA
|
| Treatment protocol |
Eight-week-old specific pathogen free Large-White piglets were infected intravenously with 10^6 copies of HEV RNA or mock-infected with PBS. At the peak of excretion (8 days post-infection), liver tissues were snap-frozen in liquid nitrogen. Samples from 2 control pigs, 3 pigs infected with HEV-3c and 3 pigs infected with HEV-3f were analyzed in this study.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
25 mg of liver was added to RLT buffer (Qiagen) supplemented with 1% β-mercaptoethanol and lysed using a Fast Prep 24 System (MP Biomedicals, Illkirch, France) in Lysis Matrix D tubes (MPBiomedicals, Illkirch, France). Total RNA was then extracted using the RNeasy minikit (Qiagen) including a digestion step on column with DNase I (Qiagen) according to the supplier’s protocol.
|
| Label |
SYBR Green
|
| Label protocol |
PCR assays were performed using the (Qiagen) following the Manufacturer’s instructions. Reverse transcription was performed from 500 ng total RNA using the RT2 First Strand Kit (Qiagen). Quantitative real-time PCR were performed (Light Cycler 96, Roche) with 45 cycles at 95 oC for 15 seconds and 60 oC for 30 seconds.
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
Test 1
|
| Data processing |
The normalization and all the data analysis were performed according to the manufacturers instructions using their web-based software package "GeneGlobe Data Analysis Center" : https://geneglobe.qiagen.com/fr/analyze
For the normalization it uses the average of four housekeeping genes: ACTG, GAPDH, PGK1 and RPL13A
Target gene signals normalized to housekeeping genes; 2^-deltaCt, where deltaCt = (Ct_Target − Ave Ct_HKG)]
The web-based software package automatically performs all deltadeltaCt based fold-change calculations from the uploaded raw threshold cycle data.
Matrix normalized worksheet reports normalized signal (against housekeeping genes).
Fold Change worksheet reports test/control (i.e., HEV-3f/Mock) ratios.
|
| |
|
| Submission date |
Sep 22, 2023 |
| Last update date |
Nov 08, 2023 |
| Contact name |
Virginie Doceul |
| E-mail(s) |
virginie.doceul@vet-alfort.fr
|
| Organization name |
UMR VIROLOGIE INRAE-ANSES-ENVA
|
| Street address |
7 avenue du Général de Gaulle
|
| City |
Maisons-Alfort |
| ZIP/Postal code |
94700 |
| Country |
France |
| |
|
| Platform ID |
GPL33783 |
| Series (2) |
| GSE243862 |
Real-time quantitative PCR analysis of liver tissue from pigs infected experimentally with hepatitis E virus (HEV) [pig liver] |
| GSE243864 |
Real-time quantitative PCR analysis of liver tissue from pigs infected experimentally with hepatitis E virus (HEV) |
|
