RIP-Seq of m6A RNA in isolated mitochondria from Brassica oleracea inflorescence rep.2
Preparation of mitochondria from cauliflower inflorescence followed the protocol described in detail in (Neuwirt et al., 2005).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.