The S. cerevisiae m6A reader Pho92 promotes timely meiotic recombination by controlling key methylated transcripts [CRAC]

We performed CRAC (UV-crosslinking and analysis of cDNA) to identify Ime4-dependent mRNA targets of Pho92 during meiosis at 5 hr in SPO. Overall design: Pho92 footprint on its RNA targets were obtained after exposing the cells to UV irradiation and purification of HTP-tagged Pho92 under stringent and denaturing conditions. Control samples includes cells expressing Pho92-HTP in ime4 mutant, Pho92-HTP cells with no UV irradiation and WT cells with no HTP-tag. Two biological replicates were performed for each sample and normalization between samples was achieved by addition, to each sample during the procedure, of an equal amount of UV-crosslinked cells of schyzosaccharomyces pombe expressing a HTP-tagged Luc7.