RNase R Digestion of Skeletal Muscle RNA

The RNase R treatment was performed essentially as described in a previous report [25 (link)]. The purified RNase R enzyme and RNase R buffer were obtained from Epicentre (Epicentre, San Diego, CA, USA). The reaction mixtures for the RNase R treatment contained 4 µg of human skeletal muscle total RNA with or without 40 units of RNase R in 40 µL solutions. The incubation for RNA digestion was performed for 30 min at 37 °C. The samples were subjected to phenol/chloroform extraction, followed by ethanol precipitation. After dissolving the precipitates in water, the nondigested human skeletal muscle total RNA (4 µg) and the RNase R-digested RNA from the same source (4 µg) were used for cDNA synthesis as described above.