2025-10-11
ENA first public
ERC000011
ENA-CHECKLIST
Viral Genetics and Biosafety Unit, French Agency for Food, Environmental and Occupational Health and Safety (ANSES), Ploufragan-Plouzané-Niort Laboratory
INSDC center name
public
INSDC status
ERS24422633
SRA accession
E-MTAB-15113:P-8933-Pu
Submitter Id
5
week
age
ArrayExpress
broker name
Not applicable
cell line
Not applicable
cell type
not collected
collection date
pig
common name
Protocols: Each experimental group of Specific Pathogen-Free (SPF) piglets was housed in a separate room in a ABSL3 facility. After 24hpi and 48hpi, different organs from the piglets, including the jejunum, duodenum, ileum, colon, lung, trachea, bronchi, trachea-bronchial and mesenteric lymph node, were harvested and placed in RNAlatter (ThermoFisher) to preserve RNA quality. Specific Pathogen-Free (SPF) piglets were oro-nasally inoculated with 7 mL of EMEM (2 mL nasal and 5mL in the oral route) for the control piglets or with 10^4.75 TCID50 per piglet, corresponding to 10^8 copies of viral N gene per pig for the infected piglets. The total RNA of jejunum crypts, 3D organoids, 2D organoids, piglet jejunums and ST cells were extracted with Trizol Reagent, according to the supplier’s recommendations. After Turbo DNAse treatment and validation of RNA quality, 500ng of RNA were used to build the sequencing librairies. Libraries were generated with the Illumina Stranded mRNA Kit, according to the supplier’s recommendations. Essentially, the mRNA were first enriched by the capture of polyA tails via oligoDT. After enzymatic fragmentation, the cDNA were synthetized and the protruding ends of the cDNA were converted into blunt ends by exonuclease/polymerase activities. The 3' ends were adenilated and the adaptators were ligated. The libraries were amplified by PCR reactions and sequenced.
description
Piglet
developmental stage
TGEV infected
disease
Not applicable
genotype
not collected
geographic location (country and/or sea)
Piglet donor 2
individual
TGEV infected
infect
10^4.75 TCID50
infection dose
not applicable
isolate
Sus scrofa
organism
Sus scrofa
organism
Intestinal jejunum
organism part
Replicate 2/3
replicate
TGEV infected piglet jejunum (10^4.75 TCID50 ; 24hpi ; piglet donor 2)
sample title
Sus scrofa
scientific_name
Female
sex
SPF Large White
strain
P-8933-Pu
title