New England Biolabs, M0288S, RNase HII

Related Categories RNases,, Epitranscriptome Analysis Specification Unit Definition One unit is defined as the amount of enzyme required to yield a fluorescence signal consistent with the nicking of 100 picomol of synthetic double-stranded DNA substrate containing a single ribonucleotide near the quencher of a fluorophore/quencher pair in 30 minutes at 37°C in 1X ThermoPol Buffer. Reaction Conditions 1X ThermoPol® Reaction Buffer Incubate at 37°C 1X ThermoPol® Reaction Buffer 20 mM Tris-HCl 10 mM (NH4)2SO4 10 mM KCl 2 mM MgSO4 0.1% Triton® X-100 (pH 8.8 @ 25°C) Storage Buffer 20 mM Tris-HCl 100 mM NaCl 1 mM DTT 1 mM EDTA 50% Glycerol Heat Inactivation No Unit Assay Conditions 1X ThermoPol Reaction Buffer with 30 nM of a synthetic dsDNA 26-mer containing an internal ribonucleotide in a total reaction volume of 150 μl. FAQ Q: RNase HII cannot be heat-inactivated. How can RNase HII be inactivated? A: RNase HII can be inactivated by adding to the reaction SDS to a final concentration of 0.1% Q: Which side of the ribonucleotide does RNase HII cut? A: RNase HII nicks 5’ to a ribonucleotide within the context of a DNA duplex. It leaves a 5’ phosphate and a 3’ hydroxyl end.