2024-03-31
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA114532194
External Id
The University of New England and CSIRO Agriculture and Food
INSDC center name
2023-10-25T14:23:18Z
INSDC last update
public
INSDC status
ERS16513526
SRA accession
E-MTAB-13418:Sample 12
Submitter Id
18
month
age
Black Baldy
breed
ArrayExpress
broker name
mononuclear cell
cell_type
neutered (steer)
clinical information
not collected
collection date
cattle
common name
Protocols: Blood samples were collected via jugular venepuncture into sterile EDTA vacutainers. Peripheral blood mononuclear cells were isolated using a gradient and resuspended in culture media before treatment. Bovine PBMCs were cultured in complete media (high glucose DMEM, 10% foetal calf serum, 1% Penicillin/Streptomycin, 1% glutamax; Thermofisher Scientific, USA) and seeded at a concentration of 10^6 cells/mL per well. Plates/samples were incubated in an incubator at 37 degrees Celcius, 5% CO2:95% air atmosphere (InCU Safe Sanyo CO2 Incubator, USA). PBMCs in samples 1-12 were treated with media only (NT), 2ug, 5ug, 12.5ug or 31.25ug Amplimune (stimulant). RNA was extracted at 0 h, 6 h or 24 h. PBMCs in samples 13-36 were treated with media only (NT) or 31.25ug Amplimune. RNA was extracted at 0 h, 6 h or 24 h. Total RNA was stored in RNAse free water in RNAse free snap-lock tubes and sent to the AGRF (Melbourne Australia) at a concentration of 1200ng. Cells were washed twice and lysed using a lysis buffer from the Isolate II RNA Mini Kit (BioLine, Meridian LifeScience Inc, USA). The lysed cells and lysis buffer were then transferred to the shredder column supplied by the kit and the protocol of the kit was used to extract RNA as per manufacturer's instructions. Ribosomal RNA depletion was done using the Ribo-Zero Gold rRNA depletion method and 36 libraries were prepared from rRNA depleted RNA.
description
adult
developmental stage
normal
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
A
individual
not applicable
isolate
Bos taurus
organism
Bos taurus
organism
blood
organism part
Bos taurus
scientific_name
male
sex
Amplimune
stimulus
Sample 12
title