Comment[ArrayExpressAccession]	E-MTAB-1608
MAGE-TAB Version	1.1																											
Investigation Title	Heat Hardening																											
Comment[Submitted Name]	Heat Hardening
Experiment Description	A common measure of heat resistance in Drosophila is the time it takes a fly to be knocked down by higher temperatures measured in a long knockdown tube or in small glass vials. This heat knockdown process involves gradual paralysis and non-responsiveness to stimuli, usually within half hour duration of constant heat exposure, and is ostensibly reversible upon return to lower temperatures, although there may be long-term fitness effects. Hardening for increased heat resistance requires exposure periods of up to 3 hours at temperatures ranging from 34C to 38C.  The experiment was aimed at understanding the heat hardening process and samples were taken during the hardening period (15 minutes, 45 minutes, 1½ hours, 2¼  hours and 3 hours).																											
Experimental Design	stimulus or stress design	time series design	in_vivo_design	replicate design	co-expression_design	growth condition design																						
Comment[AEExperimentType]	transcription profiling by array																											
Comment[AEExperimentDisplayName]	Transcription profiling by array of Drosophila melanogaster male flies to identify transcriptome changes during the heat hardening process																											
Comment[AdditionalFile:GZ]	drosophila2dmenstcdf_15.0.0.tar.gz																											
Experimental Factor Name	time	temperature																										
Experimental Factor Type	time	temperature																										
Public Release Date	2013-08-01																											
Person Last Name	Schuster																											
Person First Name	Eugene																											
Person Mid Initials	F																											
Person Email	e.schuster@ucl.ac.uk																											
Person Phone																												
Person Address	Darwin Building, Gower St., London, WC1e 6BT, UK																											
Person Affiliation	Institute of Healthy Ageing, Research Dept. GEE, UCL																											
Person Roles	submitter																											
PubMed ID																												
Publication Author List																												
Publication Title																												
Publication Status																												
Protocol Name	P-MTAB-32134	P-MTAB-32135	P-MTAB-32136	P-MTAB-32137	P-MTAB-32138
Protocol Type	growth protocol	treatment protocol	nucleic acid extraction protocol	nucleic acid labeling protocol	normalization data transformation protocol																							
Protocol Description	Flies were grown in at 18C until adulthood and then placed in cryovials and maintained in a 18C or 36C incubator. No CO2 was used during the sampling as this has been observed to have a significant effect on heat hardening.	Control flies were maintained in a 18C incubator and heat harderned flies were placed in a 36C incubator.	Briefly, samples were homogenised in 1 ml TRIzol in FastPrep tubes (Lysing Matrix D; Q-Biogene, Morgan Irvine, CA, USA) using a bead mill (Hybaid RiboLyser; Hybaid, Teddington, UK). Total RNA was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) following the manufacturers instructions. 	The RNA was further processed into cRNA using standard Affymetrix protocols.	Probes from the Affymetrix Drosophila_2 Gene Chips were mapped to up-to-date Ensembl transcript annotation with Custom CDF Drosphila2_Dm_ENST. To remove background and probe sequence biases and to summarize probes into probeset values we used gcrma in R version 2.15. Probesets that could be mapped to mRNA transcripts that were expressed were retained for loess probeset-level normalization.  If a probeset mapped to more than one transcript, then only one transcript was used for subsequent analysis.																							
Protocol Term Source REF	EFO	EFO	EFO	EFO	EFO																							
Term Source Name	EFO	ArrayExpress
Term Source File	http://www.ebi.ac.uk/efo	http://www.ebi.ac.uk/arrayexpress																										
SDRF File	E-MTAB-1608.sdrf.txt