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To protect your future RNA experiments, take the following precautions: Use our designated RNase pipettes to handle RNAse A solutions Use filter tips to handle RNase A stock solution Spin tubes containing RNase A solutions before opening Discard RNAse A waste and contaminated plasticware directly into waste bin - not your tip waste or a beaker that may be reused Treat RNase A stock like it is toxic even if it isn’t Resuspend 1-10 million yeast cells in 300 µl PBST, add 4 µl 20 mg/ml RNase A Incubate overnight at 37°C Pulse to 10k to pellet cells, remove RNase A solution Wash cells with 300 µl PBST Resuspend cells in 100-1000 µl of PBST (depending on how many cells stained) + 1 µM SYTOX (from 5 mM stock in DMSO, so 1:5,000) Let cells stain overnight at 4°C or for 1 hour at room temperature Run directly on flow * NOTE: If the samples are going to run on the ImageStream, this protocol will generate overstained samples. 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