GSM2059553: mRNA-control; Drosophila melanogaster; RNA-Seq
Approximately 180 adult female flies were used per sample. Flies were transferred to a 50 ml conical tube and snap frozen in liquid nitrogen. Heads were snapped off from the body by briefly vortexing the tubes. The frozen heads were sorted from the bodies using a series of cooled metal mesh sieves. Total RNA was isolated from the heads using the guanidinium thiocyanate single-step method (Ausubel, 1994). After isolation, RNA was treated with DNase I (Life Technologies, Grand Island, NY) and purified by acid-phenol/chloroform extraction and ethanol precipitation. RNA concentration and quality was assessed using a 2100 Bioanalyzer RNA 6000 Pico Chip (Agilent Technologies, Inc., Santa Clara, CA). RIN values for the control and ethanol-treated samples were 6.80 and 6.40, respectively. Poly(A)+ RNA was prepared from an aliquot of each total RNA sample with magnetic oligo-(dT) beads (Dynabeads® Oligo (dT), Life Technologies, Grand Island, NY). cDNA synthesis and Illumina library construction were performed with the TruSeq RNA Library Preparation Kit using standard Illumina protocols
GEO Accession
GSM2059553