Comment[ArrayExpressAccession]	E-MTAB-1035				
Investigation Title	Ontogeny of erythroid lineage differentiation				
Comment[Submitted Name]	Ontogeny of erythroid lineage differentiation				
Experiment Description	Delineate gene anatomy of differentiating primary primitive and definitive RBCs				
Comment[AEExperimentDisplayName]	Transcription profiling by array of differentiating primary primitive and definitive mouse red blood cells				
Comment[AEExperimentType]	transcription profiling by array				
Experimental Design	development_or_differentiation_design	cell_type_comparison_design			
Experimental Factor Name	phenotype	cell type			
Experimental Factor Type	phenotype	cell type			
Experimental Factor Term Source REF	MGED Ontology	MGED Ontology			
Person Last Name	Palis	Kingsley	Stoeckert	Wang	
Person First Name	James	Paul	Chris	Qi	
Person Email		paul_kingsley@urmc.rochester.edu	stoeckrt@pcbi.upenn.edu		
Person Phone		585-275-5852	215-573-4409		
Person Address		Box 703, 601 Elmwood Ave., Rochester, NY USA 14642	1415 Blockley Hall, Center for Bioinformatics, 423 Guardian Dr., Philadelphia, PA 19104		
Person Affiliation	University of Rochester	University of Rochester	University of Pennsylvania	Rutgers	
Person Roles	investigator	submitter	investigator	investigator	
Date of Experiment	2008-05-22				
Public Release Date	2012-11-21				
Protocol Name	P-MTAB-25894	P-MTAB-25895	P-MTAB-25896	P-MTAB-25897	P-MTAB-25898
Protocol Description	All samples were isolated using FACS. Because primitive erythroid cells mature semi-synchronously in the circulation, we used time as a component for their isolation: 1) proerythroblasts were isolated from e9.5, 2) basophilic erythroblasts were isolated from e10.5, 3) mix of orthochromatic and polychromatophilic erythroblasts were isolated from e12.5, 4) reticulocytes were isolated from e15.5, using scatter characteristics to purify them from co-occurring definitive fetal reticulocytes. Purity was confirmed using globin specific PCR. Fetal definitive erythroid cells are produced asynchronously and cells were isolated from e14.5, except for reticulocytes isolated from e15.5. Bone marrow derived adult erythroid stages were isolated from mature females.	RNA extraction using  the Qiagen RNeasy Plus mini kit and gDNA columns (CAT# 74134)  including Qiashredder (CAT# 79654) as described in the manufacturer's protocol.	Labeling with NuGen ovation Whole Blood Reagent according to manufacturer's protocol and recommendations.	The fragmented cRNA is mixed with the hybridization cocktail, and heat shocked at 99C for 5 minutes. In the meanwhile, the probe array is filled with hybridization buffer and incubated at 45C for 10 minutes with rotation. The heated hybridization cocktail is then transferred to a 45C heat block for 5 minutes, and spun at maximum speed in a microcentrifuge for 5 minutes. Fill the probe array cartridge with the clarified hybridization cocktail and place probe array in rotisserie box in 45C oven. Hybridize for 16 hours.	gcRMA quantification (bioconductor)
Protocol Parameters					R version;bgversion;fast;gcrma version;normalize;optical.correct;rho;type
Protocol Type	fractionate	nucleic_acid_extraction	labeling	hybridization	bioassay_data_transformation
Term Source Name	MGED Ontology				
Term Source File	http://mged.sourceforge.net/ontologies/MGEDontology.php				
Term Source Version	1.3.1.1				
SDRF File	E-MTAB-1035.sdrf.txt				
Publication DOI	 10.1182/blood-2012-04-422394				
PubMed ID	23243273				
Publication Author List	Kingsley PD1, Greenfest-Allen E, Frame JM, Bushnell TP, Malik J, McGrath KE, Stoeckert CJ, Palis J				
Publication Title	Ontogeny of erythroid gene expression