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.
- Stable and active under a wide variety of reaction conditions
- Can be heat inactivated in 30 minutes at 100°C
- Removal of RNA from DNA solutions (see Reference 2)
- Removal of RNA from recombinant protein preparations
- Ribonuclease protection assays (see Reference 3)
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.
|Definition of Activity Unit||One 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.|
|Inactivation||Inactivated by heating at 100°C for 30 min, reliably removed by spin column or phenol/chloroform extraction.|
|Inhibition||Inhibitors: SDS at 0.1% concentration irreversibly inactivates the enzyme.|
|Molecular Weight||29.6 kDa monomer|
|Quality Control||The absence of endo- and exodeoxyribonucleases confirmed by appropriate quality tests.|
|Source||E. coli cells with a cloned rna gene of E.coli.|
|Storage Buffer||The 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.|
|Storage Condition||-20 C|
|Storage Condition:||-20 C|
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