GSM4563316: Agnostic(ts3)_heat_shocked_rep1; Drosophila melanogaster; miRNA-Seq
Total RNA was isolated using Tri-reagent (Sigma-Aldrich, USA). A total of 25 µg of total RNA were separated using 15% polyacrylamide gel electrophoresis containing 8 M of urea. After incubation in ethidium bromide solution (0.5 µg/ml), gel fragments corresponding to small RNAs fraction were excised. We used chemically synthesized RNAs (Syntol, Russia) corresponding to 21 and 27 nucleotides as size markers. Cloning of small RNA libraries was performed by Illumina TruSeq Small RNA prep kit (Illumina, USA) according to the manufacturer's protocol.