GSM9833610: Isolated Male 1; Drosophila melanogaster; RNA-Seq
For each sample, flies were decapitated on dry ice. RNA was extracted and purified from homogenized whole heads using a phenol chloroform organic extraction. Samples were also treated using either the RQ1 RNase-Free DNase kit (Promega Corporation) or the TURBO DNA-free Kit (Invitrogen) to address genomic DNA contamination. The samples submitted for RNA-seq were extracted from 200 whole heads each. RNA-seq was performed by the Princeton University Genomics Core Facility. The integrity of total RNA samples was assessed on the 2100 Bioanalyzer system using the RNA 6000 Pico Chip (Agilent Technologies, CA). The poly-A containing RNA transcripts were enriched from one microgram of total RNA for each sample using oligo-dT beads, further fragmented, and converted to cDNA and Illumina sequencing libraries using the PrepX RNA-seq library preparation protocol on the Apollo 324 NGS Library Prep System (Takara Bio, CA). Different DNA barcodes were attached to each library. The RNA-seq libraries were examined on Agilent Bioanalyzer DNA High Sensitivity chips for size distribution, quantified by Qubit fluorometer (Invitrogen, CA), and pooled at equal molar amounts. Each library pool was denatured and sequenced on one Illumina NovaSeq 6000 S Prime lane using the 100 cycle v1.5 kit. Raw sequencing reads were filtered by Illumina NovaSeq Control software, and only the Pass-Filter (PF) reads were used for further analysis.
GEO
GSM9833610_r1