GSM2359512: miRNA-EtOH_30min; Drosophila melanogaster; miRNA-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. Small RNAs (18-30 nuclotides) were isolated by PAGE size fractionation.Small RNA libraries were generated from each RNA sample using Illumina Small RNA Sample Preparation kit according to the manufacturer’s instructions and sequenced with the Illumina HiSeq 2000 at the Beijing Genomics Institute (Shenzhen, China).
GEO Accession
GSM2359512