GSM8721512: BynRas1; Drosophila melanogaster; RNA-Seq
Hindgut dissection and RNA extraction using Triazol reagent QuantSeq-LEXOGEN (3' mRNA-seq Library Prep Kit for Illumina) RNA quality was assessed on a Bioanalyzer (Agilent Technologies) with RNA 6000 Nano Kit reagents and protocol (Agilent Technologies), followed by use of the 3′ mRNA-Seq Library Prep Kit protocol for Ion Torrent (QuantSeq-LEXOGEN Vienna, Austria) according to manufacturer's instructions. Briefly, up to 500 ng of RNA was used for first and second-strand synthesis, followed by amplification. Library quality and quantity were assessed on a Bioanalyzer with DNA High Sensitivity Kit reagents and protocol (Agilent Technologies). The libraries were pooled together and templated and enriched on an Ion Proton One Touch system. Templating was performed using the Ion PI Hi-Q OT-II 200 Kit (ThermoFisher Scientific), followed by sequencing with the Ion PI Hi-Q Sequencing 200 Kit on Ion Proton PI V2 chips (ThermoFisher Scientific) on an Ion Proton System, according to the manufacturer's instructions.