title: GSM8740706: chm_1_Starved_03042020_lane4_2; Drosophila melanogaster; RNA-Seq
library_construction_protocol: 30 heads were homogenized with an electrical stirrer with 500 μl of Trizol (Thermo Fisher; cat. no. 15596026). Chloroform was added at the ratio of 1:5 with Trizol and the solutions were mixed for 15 s by inverting the eppendorfs. After 5 min incubation at room temperature, samples were centrifuged at 12,000 x g for 15 min. Aqueous phase was transferred to a new eppendorf from the centrifuged sample, to which isopropanol was added at 1:1 ratio of the obtained aqueous phase, vortexed briefly and incubated for 10 min at room temperature. They were then centrifuged for 10 min at 12,000 x g. Supernatant was discarded and obtained pellet was washed with 750 μl of 80% ethanol. After brief vortexing, samples were centrifuged at 8,000 x g for 5 min. Obtained supernatant was discarded and pellet was air dried for 5 min inside the hood and resuspended in RNase-free water 1 ug of RNA, obtained from fly heads was used for library preparation. Both total and mRNA quality was assessed on a 2100 Bioanalyzer (Agilent Technologies, Cat. No G2939BA) using RNA pico assay kit (Agilent RNA 6000 Pico Kit, cat. No: 5067-1513) using manufacturer's protocol. rRNA depletion was performed using NEBNext rRNA Depletion Kit (Human/Mouse/Rat) (NEB #E6310) and library preparation for RNA-sequencing was performed using NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (NEB #E7760) following manufacturer's protocol.
identifiers.namespace: GEO
identifiers.id: GSM8740706_r1