Contr_48h_1
Anopheles gambiae
biological replicate Protocols: Larvae were reared at low density in pans with de-ionized water and fed daily with ground Tetramin fish food. Adult mosquitoes were kept in the insectary conditions at 27 C and 12h light/12h dark cycle, with constant access to 10% sucrose solution. If not otherwise stated in the treatment section, the adults were kept at 80% relative humidity. Samples were ground in TRIzol and stored at -70 C until RNA isolation. Total RNA was isolated using PureLink RNA Mini Kit (Ambion) with an on-column DNase treatment step according to the manufacturer's protocol. The libraries were constructed with the PerkinElmer Sciclone NGS Workstation using the TruSeq RNA protocol (Illumina). The mRNA was isolated from 1 ug of total RNA by a poly-A+ pull-down using biotin beads, and fragmented. Following first- and second-strand synthesis, the cDNA fragments were blunt-ended, and then their 3 prime ends were adenylated to prevent formation of chimeric molecules during the adapter ligation. Then, adapters were ligated and the fragments were subjected to a bead-based size selection using Beckman Coulter XP beads (Beckman Coulter). Prior to hybridisation to the flow cell, the cDNA constructs were amplified by PCR to selectively enrich fragments with adapter molecules on both ends. The concentration of the libraries was determined by using a Qubit dsDNA HS Assay (Life Technologies) and qPCR.
ENA-FIRST-PUBLIC
2015-07-07T17:01:16Z
ENA-LAST-UPDATE
2018-03-08T21:38:31Z
External Id
SAMEA2733680
INSDC center name
The Pirbright Institute
INSDC first public
2015-07-07T17:01:16Z
INSDC last update
2018-03-08T21:38:31Z
INSDC status
public
Submitter Id
E-MTAB-2877:Contr_48h_1
age
mock exposed to untreated test paper when 3-5 days old, culled 48 h post-exposure
broker name
ArrayExpress
common name
African malaria mosquito
developmental stage
adult
environmental history
Colonized mosquitoes derived from an eastern Ugandan population, known to have extremely high physiological resistance to pyrethroids
sample name
E-MTAB-2877:Contr_48h_1
scientific_name
Anopheles gambiae
sex
female
specimen with known storage state
fresh specimen