SAMN33829273
External Id
INRAE
INSDC center name
2023-09-17T00:50:22.807Z
INSDC first public
2023-09-17T00:50:22.807Z
INSDC last update
SRS17245479
INSDC secondary accession
live
INSDC status
Model organism or animal
NCBI submission model
Model.organism.animal.1.0
NCBI submission package
SRS17245479
SRA accession
~5 day-old pupae
age
INRAE DGIMI
biomaterial_provider
Calyx cells dissected from dsU16-injected Hyposoter didymator females at pupal stage 3 for pupae with pigmented thorax and abdomen (~5 day-old pupae), using three biological replicas (each corresponding to 30 to 50 calyx). The gene-specific double-stranded RNA (dsRNA) used for RNAi experiments were prepared with the T7 RiboMAX Express RNAi System (Promega) according to the manufacturers instructions. Briefly, a 600-750 bp fragment corresponding to U16 sequence was cloned into the double T7 vector L4440. Then, a transcription in vitro template DNA was PCR amplified with T7 primer and used to synthetized sense and antisense RNA strands with T7 RNA polymerase at 37C during 5 h. After annealing and DNase treatment with the TURBO DNA-free Kit (Life Technologies), purified dsRNA were re-suspended in nuclease-free water, quantified using a NanoDrop ND-1000 Spectrophotometer (Thermo Scientific) and examined by agarose gel electrophoresis to ensure their integrity. Injections were performed in less than one-day old female pupae using a microinjector Fentojet Express (Eppendorf) and a micromanipulator (Narishige). Approximately 0.3-0.6 ul of 2-4 ug/ul dsRNA was injected into each individual. Treated pupae were kept in an incubator until adult emergence, which occurred approximately 5 days after injection.
description
pupal stage 3 = pupae with pigmented thorax and abdomen (~5 day-old pupae)
dev_stage
Hyposoter didymator
http://purl.obolibrary.org/obo/NCBITaxon_260305
organism
female
sex
DGIMI
strain
calyx
tissue
dsU16-injected Hyposoter didymator calyx cells at pupal stage 3
title
less than one-day old female pupae were injected with U16 gene -specific double-stranded RNA (dsRNA)
treatment