New England Biolabs, M0297L, RNase H

RNase H has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10250970. Related Categories RNases,, Epitranscriptome Analysis,, cDNA Synthesis & Reverse Transcriptases Applications RT-PCR & cDNA Synthesis,, PCR Specification Unit Definition One unit is defined as the amount of enzyme required to produce 1 nmol of ribonucleotides from 20 picomoles of a fluorescently labelled 50 base pair RNA-DNA hybrid in a total reaction volume of 50 μl in 20 minutes at 37°C. Reaction Conditions 1X RNase H Reaction Buffer Incubate at 37°C 1X RNase H Reaction Buffer 50 mM Tris-HCl 75 mM KCl 3 mM MgCl2 10 mM DTT (pH 8.3 @ 25°C) Storage Buffer 50 mM KCl 10 mM Tris-HCl 0.1 mM EDTA 1 mM DTT 200 µg/ml Recombinant Albumin 50% Glycerol pH 7.4 @ 25°C Heat Inactivation 65°C for 20 minutes FAQ Q: When I thaw the 10X RNase H Reaction Buffer, I see a white precipitate.  What is it and what should I do about it? A: The white precipitate is DTT in the 10X buffer, and it will resuspend when thawed and mixed. The resuspended buffer is fully functional for its intended use. Q: Is DNA Polymerase I active in RNase H buffer? A: Yes. DNA Polymerase I is active in RNase H buffer. However, for the use of both enzymes in one reaction the use of NEBuffer 2 is recommended. Q: Which side of the RNA base does RNase H cut? A: RNase H cleaves the 3'-O-P-bond of RNA. It generates 3' hydroxyl and 5' phosphate products. Q: What is the activity of RNase H at 30°C? A: The activity of RNase H at 30°C is ~90%.