Comment[ArrayExpressAccession]	E-MTAB-3264
MAGE-TAB Version	1.1				
Investigation Title	Transcriptome of developing testes from the African malaria mosquito Anopheles gambiae				
Comment[Submitted Name]	Transcriptome of developing testes from the African malaria mosquito Anopheles gambiae
Experiment Description	Spermatogenesis is a highly complex developmental process, during which diploid germline stem cells are transformed into motile haploid spermatozoa. The process involves a precisely regulated action of a large number of genes, making the testes the most distinct tissue within the body. Testis transcriptome has been analysed in several groups of animals, but never systematically studied in mosquitoes. This dataset, consisting of the transcriptome of the developing testes from late larvae and early pupae of the African malaria mosquito, Anopheles gambiae, closes the existing gap.				
Experimental Design	development or differentiation design				
Experimental Design Term Source REF	EFO				
Experimental Design Term Accession Number	EFO_0001746				
Experimental Factor Name	replicate				
Experimental Factor Type	replicate				
Experimental Factor Term Source REF					
Experimental Factor Term Accession Number					
Person Last Name	Krzywinski				
Person First Name	Jaroslaw				
Person Mid Initials					
Person Email	jaroslaw.krzywinski@pirbright.ac.uk				
Person Phone					
Person Fax					
Person Address	Ash Road Pirbright UK				
Person Affiliation	The Pirbright Institute				
Person Roles	submitter				
Date of Experiment	2014-10-22				
Public Release Date	2016-01-21				
PubMed ID					
Publication DOI					
Publication Author List	Graham Rose, Elzbieta Krzywinska, Jan Kim, Loic Revuelta, Jaroslaw Krzywinski				
Publication Title	Dosage compensation of the X chromosome in the African malaria mosquito Anopheles gambiae				
Publication Status	submitted				
Publication Status Term Source REF	EFO				
Publication Status Term Accession Number	EFO_0001794				
Protocol Name	P-MTAB-43007	P-MTAB-43008	P-MTAB-43009	P-MTAB-43010	P-MTAB-43011
Protocol Type	growth protocol	dissection protocol	nucleic acid extraction protocol	nucleic acid library construction protocol	nucleic acid sequencing protocol
Protocol Term Source REF	EFO	EFO	EFO	EFO	EFO
Protocol Term Accession Number	EFO_0003789	EFO_0005519	EFO_0002944	EFO_0004184	EFO_0004170
Protocol Description	An. gambiae G3 strain mosquitoes used in the study were kept in the insectary conditions at 27 °C, 80% RH, and 12h light/12h dark cycle. Larvae were reared in pans with de-ionized water and fed daily with ground fish food (Tetramin). Pans containing fully grown 4th instar larvae were inspected every hour and newly pupated individuals were transferred into clean de-ionized water, and dissected within four hours after collection. Similarly, 3rd and early 4th instar larvae were collected, rinsed in clean water, and used for dissection. 	Testes were dissected under a stereomicroscope using sharpened entomological pins size 000. Care was taken to collect only the gonads with a fragment of vas efferens, but occasionally clumps of fat body cells tightly attached to the apical part of the testis were also taken. In total, testes were dissected from approximately 200 3rd instar and early 4th instar larvae and from 2000 early (up to 4 h old) pupae.  	Dissected tissues were transferred immediately into RNAlater (Ambion) and stored at -20 °C until RNA isolation. Total RNA was isolated using RNeasy Mini kit (Qiagen) and treated with TURBO DNase (Ambion) according to manufacturers’ protocols.  Then, polyA+ mRNA was isolated using Oligotex mRNA Mini kit (Qiagen) and its integrity evaluated using the Agilent 2100 Bioanalyzer. 	The mRNA was used to synthesize double-stranded cDNA using the SMARTer™ PCR cDNA synthesis kit (Clontech) according to the manufacturer’s recommendation. Fragmentation of 1 μg of double-stranded cDNA by nebulization, fragment ends polishing, adaptor ligation, isolation and estimation of the single-stranded template library concentration, emulsion PCR with library beads, and loading on the 454 plate was performed according to the published protocol (Margulies M, et al. 2005. Nature 437: 376-380).	Pyrosequencing was conducted on the 454 GS FLX platform (Roche 454 Life Sciences) using Titanium chemistry according to the manufacturer's protocol.
Protocol Hardware					454 GS FLX
Protocol Software					
Term Source Name	EFO
Term Source File	http://www.ebi.ac.uk/efo/				
Term Source Version					
Comment[AEExperimentType]	RNA-seq of coding RNA				
Comment[SecondaryAccession]	ERP009392
Comment[SequenceDataURI]	http://www.ebi.ac.uk/ena/data/view/ERR744531-ERR744532
SDRF File	E-MTAB-3264.sdrf.txt