Spin x centrifuge tubes

Total RNA was isolated using Trizol (Thermo Fisher Scientific) from three replicates of 30 4–7 day-old female flies after desiccation stress and under control conditions. RNA samples were treated with DNAse I (Thermo Fisher Scientific) following manufacturer’s instructions. Small RNAs were obtained by gel size selection from total RNA using 15% Mini-Protean TBE-Urea Gel (Bio-Rad, #4,566,056). The gel was run at 300 V for 50 min. Fragments between 17 and 30 nucleotides were carefully cut from the gel and were put in an Eppendorf with 250 μL NaCl 0.5 M and then were placed at 4 °C in a rotating wheel o/n. Next, the sample was transferred to Corning Costar Spin-X centrifuge tubes (Merck, CLS8162), spinned and cleaned with standard ethanol washes, and eluted in DEPEC-H2O. Quality check was performed using Bioanalyzer small RNA kit (Agilent). Libraries were sequenced using Illumina Next-Seq 50 bp single-end reads.

Total RNA was isolated using Trizol (Thermo Fisher Scientific) from three replicates of 30 4–7 day-old female flies after desiccation stress and under control conditions. RNA samples were treated with DNAse I (Thermo Fisher Scientific) following manufacturer’s instructions. Small RNAs were obtained by gel size selection from total RNA using 15% Mini-Protean TBE-Urea Gel (Bio-Rad, #4,566,056). The gel was run at 300 V for 50 min. Fragments between 17 and 30 nucleotides were carefully cut from the gel and were put in an Eppendorf with 250 μL NaCl 0.5 M and then were placed at 4 °C in a rotating wheel o/n. Next, the sample was transferred to Corning Costar Spin-X centrifuge tubes (Merck, CLS8162), spinned and cleaned with standard ethanol washes, and eluted in DEPEC-H2O. Quality check was performed using Bioanalyzer small RNA kit (Agilent). Libraries were sequenced using Illumina Next-Seq 50 bp single-end reads.