<i>In vitro</i> endonuclease actictivity with different RNA substrates.

Abstract

A 10 μL reaction containing 0.5 μM full-length SNV L K124A, radiolabeled RNA substrate, and 0.1 mM (low MnCl2) or 10 mM (high) MnCl2 or MgCl2 in reaction buffer was incubated at 37°C for one hour. The reactions were stopped by adding 10 μL of 2x RNA loading dye (98% formamide, 18 mM EDTA, 0.025 mM SDS, xylene cyanol, bromophenol blue) and heated to 95°C for five minutes. RNA was separated on denaturing PAGE (25% acrylamide, 7 M urea, 0.5 x TBE) and visualized by autoradiography. The control lane shows a reaction without the addition of SNV L K124A. (TIFF)</p