2.5 μg of in vitro-transcribed RNAs (Circular EGFP ΔpAΔIRES) were incubated at 37°C with 10 U of RNase R (Cosmo Bio) in a 10 μl mixture. After a 45 min incubation, the mixture was subjected to 4 and 8% denaturing PAGE (8.3 M urea), and then stained by SYBR Green II Nucleic Acid Gel Stain (TaKaRa). Stained RNA was detected by Typhoon FLA-7000 (GE Healthcare).
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