organism: Aedes aegypti
description: Protocols: 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
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2013-09-01T17:01:51Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T16:22:13Z
attributes.tag: External Id
attributes.value: SAMEA2064058
attributes.tag: INSDC center name
attributes.value: CNRS
attributes.tag: INSDC first public
attributes.value: 2013-09-01T17:01:51Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T16:22:13Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-1635:Propo-R
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: yellow fever mosquito
attributes.tag: developmental stage
attributes.value: larval stage
attributes.tag: phenotype
attributes.value: carbamate insecticide propoxur resistant
attributes.tag: sample name
attributes.value: E-MTAB-1635:Propo-R
attributes.tag: scientific_name
attributes.value: Aedes aegypti
attributes.tag: specimen with known storage state
attributes.value: fresh specimen
attributes.tag: strain
attributes.value: Propo-R