Ribonuclease H (RNase H) is an endoribonuclease that specifically degrades the RNA strand of an RNA-DNA hybrid to produce 5´ phosphate-terminated oligoribonucleotides and single-stranded DNA. Applications Removal of mRNA during second-strand cDNA synthesis. Removal of poly(A) sequences from mRNA in the presence of oligo(dT). Oligodeoxyribonucleotide-directed cleavage of RNA. Source Purified from E. coli expressing the E. coli RNase H gene on a plasmid. Performance and quality testing Ribonuclease, nonspecific endodeoxyribonuclease, 3´ and 5´ exodeoxyribonuclease. Unit definition One unit hydrolyzes 1 nmol of RNA in 3H-labeled poly(A)poly(dT) to acid-soluble material in 20 min at 37°C. Unit reaction conditions 20 mM Tris-HCl (pH 7.5), 0.1 M KCl, 10 mM MgCl2 , 0.1 mM DTT, 5% (w/v) sucrose, 0.5 nmol 3H-labeled poly(A)poly(dT), and enzyme in 50 µL for 20 min at 37°C.
For Research Use Only. Not for use in diagnostic procedures.
Approved for shipment on Wet or Dry Ice
Contents & storage
1 vial of Ribonuclease H containing 30 units supplied at a concentration of 2U/µl