Comment[ArrayExpressAccession]	E-GEOD-46176
MAGE-TAB Version	1.1							
Public Release Date	2013-08-01
Investigation Title	Expression of circulating microRNAs in Influenza A patients.							
Comment[Submitted Name]	Expression of circulating microRNAs in Influenza A patients.
Experiment Description	Altered expression of microRNAs in circulation, in H1N1 infected patients, were determined using expression profiling microRNA expression profiling of blood samples from H1N1 infected patients							
Term Source Name	ArrayExpress	EFO
Term Source File	http://www.ebi.ac.uk/arrayexpress/	http://www.ebi.ac.uk/efo/efo.owl						
Person Last Name	Jeyaseelan	Tambyah	Sepramaniam	Ali	Ching	Swaminathan	Armugam	Jeyaseelan
Person First Name	Kandiah	Paul	Sugunavathi	Jaminah	Chai	Priyadharshini	Arunmozhiarasi	Kandiah
Person Mid Initials		A		M	S			
Person Email	bchjeya@nus.edu.sg							
Person Affiliation	National University of Singapore							
Person Phone	+6565163248							
Person Fax	+6567791453							
Person Address	Biochemistry, National University of Singapore, 8 Medical Drive, Singapore, Singapore							
Person Roles	submitter							
Protocol Name	P-GSE46176-1	P-GSE46176-4	P-GSE46176-5	P-GSE46176-2	P-GSE46176-3	P-GSE46176-6		
Protocol Description	miRNA microarray data were analyzed by subtracting the background signal (B532nm) from the test signal (F532nm). ID_REF =  VALUE = background subtracted signal value (F532 - B532)	LNA-modified oligonucleotide (Exiqon, Denmark) probes for human, mouse and rat miRNAs annotated in miRBase version 16.0 were used in the microarray. Total RNA (1 M-5g) was 3M-bM-^@M-2-end M-bM-^@M-^Slabeled with Hy3 dye using the miRCURY LNAM-bM-^DM-" Power Labeling Kit (Exiqon, Denmark).	Hybridization on miRCURY LNAM-bM-^DM-" Arrays, using MAUIM-. hybridization system.	Primary cultures of cortical neurons were established from E15 Swiss albino mouse brains. The cortices were dissected from E15 mouse embryos and washed with HanksM-bM-^@M-^Y balanced salt solution (HBSS, 14025-092, Gibco, Invitrogen, USA). The cortical slices was dissociated with 0.05% (w/v) trypsin in HBSS without Ca2+/Mg2+ (14175-095, Gibco, Invitrogen, USA) for 30 min at 37 M-0C and neutralized with 1mg/ml trypsin inhibitor (T6522, Sigma, USA). Single cells were obtained by gentle trituration in Neurobasal medium (21103-049, Gibco, Invitrogen, USA) supplemented with B27 (17504-044, Invitrogen, USA), L-glutamine and Penicillin-streptomycin (Gibco, Invitrogen, USA). The cells were counted by trypan blue exclusion and seeded on to poly-d-lysine coated 24 well plates at a density of 120, 000 cells/cm2. Cultures were maintained at 37 M-0C with 5% CO2 in a tissue culture incubator. Cells were harvested on Days 2, 4, 6, 8.	Total RNA (+ microRNA) was extracted from cells by Ribopure Kit (Ambion, USA) according to the manufacturersM-bM-^@M-^Y protocol. The concentration and integrity of the RNA were determined using Nanodrop ND-2000c spectrophotometry (Nanodrop Tech, Rockland, Del) and denaturing polyacrylamide gel electrophoresis, respectively.	Hybridization images were scanned and digitized using InnoScan 700 microarray scanner (Innopsys, France).		
Protocol Type	normalization data transformation protocol	labelling protocol	hybridization protocol	growth protocol	nucleic acid extraction protocol	array scanning protocol		
Experimental Factor Name	INFECTION STATUS							
Experimental Factor Type	infection status							
Comment[SecondaryAccession]	GSE46176							
Comment[GEOReleaseDate]	2013-08-01							
Comment[ArrayExpressSubmissionDate]	2013-04-18							
Comment[GEOLastUpdateDate]	2013-08-03							
Comment[AEExperimentType]	transcription profiling by array							
SDRF File	E-GEOD-46176.sdrf.txt