6 protocols
AccessionType
image_acquisition
Scanning was performed with a Agilent 2565 AA DNA Microarray scanner using defaults parameters (100 PMT, 10 um resolution, at 20 degrees C in low ozone concentrationenvironment. Microarray images were alysed by using Feature Extraction software version A.8.1.1 from Agilent technologies. Defaults settings were used.
hybridization
Agilent Hybridization Protocol (Chamber type: Agilent SureHyb Chamber; Quantity of labelled extract: .75 ug; duration: 17 hours; volume: 500 ul ; Temperature in °C: 60).
labeling
Probe synthesis was done with Agilents low RNA input kit,using 200ng of total RNA template, following Agilent protocol
nucleic_acid_extraction
RNA isolation was done using Qiagen RNAeasy tissue kit
infect
Animals were confirmed seronegative against current H1N1 and H3N2 influenza reference viruses by haemagglutination inhibition assay. Animals were infected with 1918 (n=7) viruses via a combination of intratracheal (4 ml), intranasal (0.5 ml/nostril), intraocular (0.5 ml/eye) and oral (1 ml) routes with suspension containing 106 PFU/ml (infectious dose by all routes = 7 x 106 PFU).
infect
Animals were confirmed seronegative against current H1N1 and H3N2 influenza reference viruses by haemagglutination inhibition assay. Animals were infected with K173 (n =3), as conventional human virus control, via a combination of intratracheal (4 ml), intranasal (0.5 ml/nostril), intraocular (0.5 ml/eye) and oral (1 ml) routes with suspension containing 106 PFU/ml (infectious dose by all routes = 7 x 106 PFU).