Chikens were inoculated with 100ul of PBS or H5N1 influenza virus intranasally.
Growth protocol
Chickens were kept in the biosafety level three facility for 24hour.
Extracted molecule
total RNA
Extraction protocol
Chicken lung soaked into RNAlater(Ambion) was homogenaized with TRIzol (Invitrogen). Total RNA was extracted using the RNeasy Mini kit (Qiagen Inc., Valencia, CA, USA) and the RNAprotect reagent (Qiagen) and DNA was removed by on-column DNase digestion with the RNase-Free DNase set (Qiagen). These processes were carried out in Nihon Gene research laboratories.
Label
Cy3
Label protocol
Labeling was performed by NimbleGen Systems Inc., Madison, WI USA. Control1, Control2, HP1 and HP2 were labelled with NimbleGen 60mer Hybriweel protocol, biotin-labeled method by in vitro transcription. For the other samples, complemental strand was synthesized from Cy3-random 9 mer using with double strand cDNA as a template.
Hybridization protocol
Hybridization was performed by NimbleGen Hybridization System, NimbleGen Systems Inc., Madison, WI, USA following their standard operating protocol. See www.nimblegen.com.
Scan protocol
Scanning was performed by Axon GenePix 4000B, NimbleGen Systems Inc., Madison, WI USA, following their standard operating protocol. See www.nimblegen.com.
Description
WB2 Gene expression of chicken lung at 24 hours post infection of influenza virus
Data processing
The raw data (.pair file) was subjected to RMA (Robust Multi-Array Analysis; Irizarry et al. Biostatistics 4(2):249), quantile normalization (Bolstad et al. Bioinformatics 19(2):185), and background correction as implemented in the NimbleScan software package, version 2.4.27 (Roche NimbleGen, Inc.).
