title: GSM4815634: D.Melanogaster_Elav-GS-Gal4>UAS-CLK-DN1_female_heads_DR-diet_Vehcle-treated_control_ZT0_Replicate-1; Drosophila melanogaster; RNA-Seq
library_construction_protocol: Flies were flash frozen on dry-ice at ZT 0 and ZT 12 (lights-on and -off, respectively). Frozen flies were vortexed to collected heads and mRNA from each biological replicated of pooled heads were isolated with the Quick-RNA MiniPrep Kit (Zymo Research #11-328), per manufacturer's protocols. Library preparation was performed by the Functional Genomics Laboratory (FGL), a QB3-Berkeley Core Research Facility at University of California, Berkeley. cDNA libraries were produced from the low-input RNA using the Takara SMART-Seq v4 Ultra-low input RNA kit. An S220 Focused-Ultrasonicator (Covaris®) was used to fragment the DNA, and library preparation was performed using the KAPA hyper prep kit for DNA (KK8504). Truncated universal stub adapters were used for ligation, and indexed primers were used during PCR amplification to complete the adapters and to enrich the libraries for adapter-ligated fragments. Samples were checked for quality on an AATI (now Agilent) Fragment Analyzer. Samples were then transferred to the Vincent J. Coates Genomics Sequencing Laboratory (GSL), another QB3-Berkeley Core Research Facility at UC Berkeley, where Illumina sequencing library molarity was measured with quantitative PCR with the Kapa Biosystems Illumina Quant qPCR Kits on a BioRad CFX Connect thermal cycler. Libraries were then pooled evenly by molarity and sequenced on an Illumina NovaSeq6000 150PE S4 flowcell, generating 25M read pairs per sample. Raw sequencing data was converted into fastq format, sample specific files using the Illumina bcl2fastq2 software on the sequencing centers local linux server system.
attributes.tag: GEO Accession
attributes.value: GSM4815634