RIP-Seq of m6A RNA in isolated mitochondria from Arabidopsis thaliana leaves rep.2
Isolation of mitochondria from 3-week-old Arabidopsis seedlings grown on MS-plates was performed according to the method described in (Keren et al., 2012). RNA was prepared following standard TRI Reagent protocols (Molecular Research Center Inc., TR-118) and was treated with RNase-free DNase (Ambion, Cat no. AM2222). m6A mthylated RNA was immunoprecipitated using Monoclonal anti-m6A antibody (Synaptic systems, Cat no. 202011) bound to Protein A/G Magnetic Beads (Pierce, 88802). Purified m6A-RNA was sequenced on an Illumina Genome Analyzer.