GSM7498808: HSD, 10days, replicate C; Drosophila melanogaster; RNA-Seq
Flies were anesthetized using FlyNap® . 15 hearts per replicate were dissected and put in 300ul TRIzol®  reagent. 30ul of RNAse-free water and 0.2ul of GenElute™ LPA (25ug/ul stock; Sigma Aldrich) were added and samples vortexed. 60ul of Chloroform was added and the volume transferred in Phase Lock Gel™  Heavy (VWR™) after vortexing. Total RNAs, contained in the aqueous phase obtained by centrifugation, were precipitated overnight at -20° in 0.8v/v 2-Propanol. After centrifugation at 4° for  30 minutes, the pellet was carefully washed in 75% EtOH, air dried and dissolved in 8.5ul RNAse-free water. Every sample was quantified using a NanoDrop™. 3 replicates for each diet were prepared for sequencing. Librairies were prepared according to the specifications of the Illumina® Stranded total RNA prep Ligation with Ribo-Zero Plus" kit .  Librairies quantity and quality were evaluated using the Agilent DNF-474 HS NGS Fragment kit according to the manufacturer's indications.