GSM3728901: rice.2.m6A.seal.ip; Oryza sativa; OTHER
Total RNA was first extracted from HeLa cells or rice tissue by Trizol reagent, The polyA RNA was then purified with oligo d(T)25 magnetic beads PolyA RNA was fragmented by NEBNext Magnesium RNA Fragmentation Module. A portion of fragmented RNA was reserved as m6A input sample.For MeRIP-m6A-seq,  the left RNA was incubated with anti-m6A antibody and then eluted to get the m6A IP sample. For m6A-SEAL, the left RNA was enzymatically oxidized by FTO and then conjugated with biotin by dithiothreitol under acidic aqueous solution. The biotinylated RNA are enriched by streptavidin beads and then eluted by dithiothreitol to get m6A IP sample. Both input and m6A IP samples were prepared for next-generation sequencing library by standard protocol of NEBNext Ultra II Directional RNA Library Prep Kit for Illumina. m6A-SEAL and MeRIP-m6A-seq. Library was sequenced in PE150 model on Illumina X-Ten platform or in SR75 model on Illumina NextSeq 500 platform.