title: GSM5466274: Testes; Drosophila melanogaster; OTHER
library_construction_protocol: A line of wild type flies Oregon R was used for transcripts analysis. Flies have been collected on 5-7 day after hatching and frozen with liquid nitrogen before heads were separated from the bodies by vigorous shaking. The eyes were separated with tweezers while the heads were frozen. Testes and heads were isolated in PBS from males on 5-7 days after hatching using small tweezers. The samples were cooled at –80°C prior to using.. tal RNA was isolated with TRI Reagent (Molecular Research Center, Inc.) according to the manufacturer's instructions. The quality of the RNAs were monitored using the Agilent 2100 Bioanalyser (Agilent) and Agilent RNA 6000 Pico Kit (Agilent). The extracted RNA was quantified with an Implen NanoPhotometer (Implen, Germany) and Qubit 4 Fluorimeter (Invitrogen). DNA impurities were removed by using DNA-free™ Kit (Invitrogen) as recommended by the manufacturer.  Total RNA was used for cDNA production. Double-stranded cDNA was obtained from total RNA by using cDNA mint kit (Evrogen) according to the manufacturer instructions. The cDNA for 3'- and 5'-terminal fragments of qtc transcripts was obtained by single rounds of amplification with the Q5 High- Fidelity 2X Master Mix (NEB, UK) by using the MINT RACE primer set (Evrogen) and specially designed gene-specific primers.