Sample 32
Gallus gallus
Protocols: Chickens were housed in 4 separate rooms (2 challenged rooms, 2 non-challenged rooms) and the two breeds were intermixed in each room in a biosafety level 2 facility. To obtain tissue samples at 2, 6, and 10 days post inoculation, approximately one-third of the birds were euthanized with sodium pentobarbital solution at each of the three time points. The lung was harvested and placed into RNAlater® solution (ThermoFisher Scientific, Waltham, Massachusetts) for short-term storage. Within a week, the lung tissue was removed from RNA-later and moved to -80°C freezer for long-term storage. Challenged chickens were inoculated with a high titered lentogenic Newcastle Disease Virus (NDV) strain via the eyes and nostrils at three weeks of age. Non-challenged chickens were given PBS via the same route. RNA was isolated from the lung tissue using the RNAqueous® kit (ThermoFisher Scientific, Waltham, Massachusetts) and samples were then DNase treated using the DNA-free™ kit (ThermoFisher Scientific, Waltham, Massachusetts). The quality of all samples was validated (RNA quality number>8.0) using the Fragment Analyzer™ (Advanced Analytical Technologies, Ankeny, Iowa). A 500 ng input for each sample was utilized to construct the cDNA library using TruSeq® RNA Sample Preparation v2 Guide (Illumina, San Diego, California) using the high throughput protocol. Following cDNA library validation using the Fragment Analyzer™ (Advanced Analytical Technologies, Ankeny, Iowa).
ENA first public
2017-07-28
ENA last update
2017-07-12
External Id
SAMEA104161553
INSDC center alias
Iowa State University
INSDC center name
Iowa State University
INSDC first public
2017-07-28T17:03:31Z
INSDC last update
2017-07-12T11:06:08Z
INSDC status
public
Submitter Id
E-MTAB-5859:Sample 32
age
31
breed
Fayoumi
broker name
ArrayExpress
common name
chicken
disease
Newcastle disease
individual
LG1576
organism part
lung
phenotype
resistant
sample name
E-MTAB-5859:Sample 32
sex
male