GSM5655659: DruB_experiments; Dictyostelium discoideum; OTHER
Cells were pelleted and resuspended in TRIzol reagent (Invitrogen) containing 10 mM EDTA (pH 8.0). RNA was extracted according to the manufacturer's instructions RiboMeth-seq: 10 ug of RNA was partially degraded by alkaline at denaturing temperatures. The size fraction 20-40 nt was purified on gels and linkers added using a system relying on a modified Arabidopsis tRNA ligase joining 2',3' cyclic phosphate and 5' phosphate ends. The library fragments were then sequenced on the Ion Proton platform. See Birkedal U, Christensen-Dalsgaard M, Krogh N, Sabarinathan R, Gorodkin J, Nielsen H. Profiling of ribose methylations in RNA by high-throughput sequencing. Angewandte Chemie. 2015;54(2):451-5 for detailed description