Comment[ArrayExpressAccession] E-MTAB-1635 MAGE-TAB Version 1.1 Investigation Title Aedes aegypti RNA seq Insecticide resistance LECA Grenoble Comment[Submitted Name] Aedes aegypti RNA seq Insecticide resistance LECA Grenoble Experiment Description RNA seq comparison of different Ae. Aegypti strains obtained by selection with various insecticides. Experimental Design strain or line design co-expression_design stimulus or stress design Comment[AEExperimentType] RNA-seq of coding RNA Comment[AEExperimentDisplayName] RNA-seq of coding RNA of different Aedes aegypti strains obtained by selection with various insecticides Experimental Factor Name strain phenotype Experimental Factor Type strain phenotype Quality Control Type biological replicate Quality Control Term Source REF EFO Public Release Date 2013-09-01 Person Last Name DAVID Person First Name Jean-Philippe Person Mid Initials Person Email jean-philippe.david@ujf-grenoble.fr Person Phone 33476514459 Person Address Laboratoire d'ecologie Alpine (LECA), 2233 Rue de la piscine, Batiment D Biologie, Campus Universitaire, 38041 Grenoble Person Affiliation CNRS Person Roles submitter PubMed ID Publication Author List Publication Title Deep analysis of adaptation to three insecticides in the dengue mosquito Aedes aegypti using RNA sequencing Mosquito resistance to Bacillus thuringiensis israelensis toxins : a whole transcriptome analysis Chemical and biological insecticides select distinct gene expression patterns in mosquito Publication Status in preparation in preparation in preparation Protocol Name P-MTAB-32415 P-MTAB-32416 P-MTAB-32417 P-MTAB-32418 P-MTAB-32419 Protocol Type growth protocol treatment protocol nucleic acid extraction protocol nucleic acid library construction protocol nucleic acid sequencing protocol Protocol Description Mosquitoes were reared in standard insectary conditions (26C, 14h/10h light/dark, 80% relative humidity) in tap water (larvae) and net cages (adults). Larvae and adults were fed with hay pellets and papers impregnated with honey respectively. Blood feeding of adult females was performed on mice. The laboratory strain Bora-Bora, originating from French Polynesia and fully susceptible to insecticides, was used as a parental strain to select different strains with the pyrethroid insecticide permethrin (Perm-R strain), the neonicotinoid insecticide imidacloprid (Imida-R strain), the carbamate insecticide propoxur (Propo-R strain) and Bacillus turingiensis toxins (LiTOX_N and LiTOX_S strains). For Perm-R, Imida-R and Propo-R strains, insecticide selection was performed by exposing early 4th-stage larvae for 24h to a lethal dose of each insecticide. For each strain, dose of insecticide was adjusted at each generation to reach 60-80% larval mortality after 24h exposure. Surviving larvae were transferred in clean tap water, fed with standard larval food and allowed to emerge. Adults were allowed to reproduce for 4-days and blood fed to obtain eggs for the next generation. In order to limit bottleneck effects, each generation was seeded with more than 6000 individuals. Selection process was carried out in parallel for all strains during 10 generations. During this process, the parental Bora-Bora strain was maintained in similar condition without insecticide selection. For LiTOX_N and LiTOX_S strains, selection as described in Paris et al. 2011 (DOI 10.1007/s10646-011-0663-8). For each strain, total RNA was extracted from G11 larvae not exposed to insecticides of three different egg batches. Each biological replicate consisted of 200 larvae reared in 200 mL tap water in standardized conditions. For each biological replicate, total RNA was extracted from 60 fourth-stage larvae using the RNAqueous-4PCR Kit (Applied Biosystems/Ambion, Austin, TX, USA), according to manufacturers instructions. Total RNA quality and quantity were assessed with a Nanodrop ND11000 (Thermo Scientific, USA) and a 2100 Bioanalyzer (Agilent, USA). After extraction, total RNAs from each biological replicate were pooled in equal quantity to obtain a total RNA mixture representative of 180 individuals. Total RNA pools from each strain were then used for preparing cDNA libraries using the mRNA-Seq Sample Prep Kit (Part 1004898 Rev D, Illumina, USA) according to manufacturers instructions. Two replicates of cDNA libraries were prepared for each strain Each cDNA library was sequenced as single reads of 75bp in a distinct flow cell lane with a Genome Analyzer II (Illumina) at the National Sequencing Center (Genoscope, Evry, France). Protocol Term Source REF EFO EFO EFO EFO EFO Protocol Hardware Illumina Genome Analyzer II Term Source Name EFO Term Source File http://www.ebi.ac.uk/efo Comment[SecondaryAccession] ERP002530 Comment[SequenceDataURI] http://www.ebi.ac.uk/ena/data/view/ERR266245-ERR266256 SDRF File E-MTAB-1635.sdrf.txt