Figure 3: hTRIR digests different forms of RNA by In vitro digestion assay.
From: Identification of an RNase that preferentially cleaves A/G nucleotides
(A) T7 transcribed α-32P-GTP labeled hTR or ampicillin resistance gene ORF (nt 1–300) was incubated with different amount of purified hTRIR in digestion buffer at 37 °C for 30 min. RNA was purified by phenol-chloroform extraction and precipitated with ethanol-glycogen, then separated by 7 M urea 6% Polyacrylamide gel electrophoresis (PAGE) at 550 V, 15 W for 90 min. Signals were detected with Typhoon Cyclone. (B) 2 μg total RNA was incubated with different amount of purified hTRIR at 37 °C for 30 min and separated by 1.5% agarose gel. Signals were detected with GelDoXR + IMAGELAB. (C) Small RNA was incubated with purified 1 μg hTRIR at 37 °C for 30 min and separated by 7 M urea 20% PAGE at 200 V, 2 W for 30 min. RNA was stained with stain-all (Sigma) and scanned with Epson scanner.
