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RNase I

LO certified Recombinant enzyme Thermal inactivation at 100°C in 30 min

Ribonuclease I (RNase I) is an endoribonuclease that hydrolyzes single-stranded RNA to nucleoside 3'-monophosphates.
  
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Thermo Scientific Ribonuclease I (RNase I), an endoribonuclease, preferentially hydrolyzes single-stranded RNA to nucleoside 3'-monophosphates via nucleoside 2', 3'-cyclic monophosphate intermediates (see Figure 1 in Supporting Data [see Reference 1]).

The enzyme does not require any metal ions for activity.

Highlights

  • Stable and active under a wide variety of reaction conditions
  • Can be heat inactivated in 30 minutes at 100°C

Applications

  • Removal of RNA from DNA solutions (see Reference 2)
  • Removal of RNA from recombinant protein preparations
  • Ribonuclease protection assays (see Reference 3)

Notes

Mammalian ribonuclease inhibitors have no effect on RNase I. RNase I binds to DNA, but does not degrade it. RNase I amino acid sequence is typical for the RNase T2 family. Non-ionic detergents (e.g., Triton X-100) do not inhibit RNase I, and may even slightly stimulate its activity and stabilize it against heat inactivation. Triton X-100 or BSA (at 0.1 mg/mL) may prevent RNase I from sticking to glass vessels when working with dilute solutions. Polyamines stimulate the activity of RNase I.

  
Storage Condition-20 C
HazardousNo
Concentration10 U/µL
Definition of Activity UnitOne unit of the enzyme catalyzes degradation of 100 ng of E. coli ribosomal RNA per second into acid-soluble nucleotides at 37°C. Enzyme activity is assayed in the following mixture: 20 mM Tris-acetate (pH 8.0), 100 mM NaCl, 0.1 mM EDTA, 0.01% (v/v) Triton X-100, 40 µM E. coli ribosomal [3H]-RNA.
InactivationInactivated by heating at 100°C for 30 min, reliably removed by spin column or phenol/chloroform extraction.
InhibitionInhibitors: SDS at 0.1% concentration irreversibly inactivates the enzyme.
Molecular Weight29.6 kDa monomer
Quality ControlThe absence of endo- and exodeoxyribonucleases confirmed by appropriate quality tests.
SourceE. coli cells with a cloned rna gene of E.coli.
Storage BufferThe enzyme is supplied in 50 mM Tris-HCl (pH 8.0), 100 mM NaCl, 0.01% (v/v) Triton X-100, and 50% (v/v) glycerol.
RNase I activity

RNase I activity

RNase I activity

RNase I activity.


References

  1. V. Shen, D. Schlessinger, RNase I of Escherichia coli, The Enzymes, Boyer, P.D., Ed. 3 (Academic Press Inc., New York, 1982), vol. 15B, pp. 503-506.
  2. L. Zhu, M. P. Deutscher, The Escherichia coli rna gene encoding RNase I: sequence and unusual promoter structure. Gene. 119, 101-106 (1992).
  3. J. Sambrook, D. W. Russell, Molecular Cloning: A. Laboratory Manual, the third edition (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2001).

MSDS & Certificates

pdf   RNase I - MSDS
pdf   MSDS (JP) - RNase I