RNase H, E. coli

Applications

  • Elimination of RNA prior to second-strand synthesis of cDNA.1
  • Removal of poly(A) tails from mRNA hybridized to oligo(dT).2
  • Specific cleavage of mRNAs after hybridization to oligonucleotide probes.3
  • Specific destruction of "hybrid-arrested" mRNAs during in vitro translation.
  • Diagnostic assays using NASBA® transcription-based amplification methods.4
  • Diagnostic assays based on Cycling Probe Technology.5
  • Template-dependent probe technologies.

Ribonuclease H (RNase H) from E. coli is an endonuclease that specifically degrades the RNA in an RNA:DNA hybrid, without affecting DNA or unhybridized RNA. It will not degrade dsDNA, ssDNA, dsRNA, or ssRNA. Epicentre's RNase H is highly purified and suitable for use in diagnostic probe research, as well as other applications. However, in contrast to Hybridase™ Thermostable RNase H, which can be used at temperatures up to 95°C, E. coli RNase H is inactivated at 65°C in 10 minutes.

Unit Definition: One unit of RNase H results in the acid-solubilization of 1 nmol of polyadenylic acid in the presence of an equimolar concentration of polythymidylic acid in 20 minutes at 37°C under standard assay conditions.

Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 M NaCl, 0.1 mM EDTA, 1 mM DTT, and 0.1% Triton® X-100.

Quality Control: RNase H is tested for the specific degradation of RNA hybridized to DNA and is free of detectable exo- and endodeoxyribonuclease, and non-RNase H RNase activities.

References

  1. Gubler, U. (1987) Meth. Enzymol. 152, 330.
  2. Vournakis, J.N. et al. (1975) Proc. Natl. Acad. Sci. USA 72, 2959.
  3. Donis-Keller, H. (1979) Nucleic Acids Res. 7, 179.
  4. Guatelli, J.C. et al. (1990) Proc. Natl. Acad. Sci. USA 87, 1874.
  5. Bekkaoui, F. et al. (1996) BioTechniques 20, 240.