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Introduction
Ribonuclease H (RNase H) is an endoribonuclease which specifically degrades the RNA strand of an RNA-DNA hybrid to produce 5' phosphateterminated oligoribonucleotides and single-stranded DNA. It is suitable for removing mRNA during second strand cDNA synthesis, removing the poly(A) sequences from mRNA in the presence of oligo(dT), and for oligodeoxyribonucleotide-directed cleavage of RNA. RNase H is purified from E. coli expressing the E. coli RNase H gene on a plasmid.
Unit Definition
One unit is defined as the amount of RNase H that solubilizes 1 nmol poly(A) in 20 minutes at +37°C
Storage Buffer
20 mM Tris-HCl (pH 7.5)
100 mM KCl
10 mM MgCl2
0.1 mM EDTA
0.1 mM DTT
50 μg/ml BSA
50% (v/v) glycerol
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