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RNase I, E.coli

#EN0601	1000 u (10 u/µl)
#EN0602	5000 u (10 u/µl)
Related Documents (in pdf, ~40-60 KB): Certificate of Analysis: #EN0601, #EN0602 MSDS (English) MSDS (English-USA) MSDS (German)

 

Feature
Stable and active under a wide variety of reaction conditions.

Description
The RNase I, E.coli, an endoribonuclease, preferentially hydrolyzes single-stranded RNA to nucleoside 3'-monophosphates via nucleoside 2'-,3'-cyclic monophosphate intermediates (1). The enzyme does not require any metal ions for activity.

Source
E.coli cells with a cloned rna gene.

Molecular Weight
29.6 kDa monomer.

Applications

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

Quality Control
The absence of endodeoxyribonucleases and exodeoxyribonucleases confirmed by appropriate quality tests.

Concentration
10 u/µl

Definition of Activity Unit
One unit of the enzyme catalyzes degradation of 100ng 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.

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.

Inhibition and Inactivation

• Inhibitors: SDS at 0.1% concentration irreversibly inactivates the enzyme.
• Inactivated by heating at 100°C for 30 min, phenol/chloroform extraction or with SDS-Proteinase K.

Note

• RNase I does not degrade DNA, but binds to it.
• Mammalian ribonuclease inhibitors have no effect on RNase I.
• 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 protect RNase I from sticking to glass vessels when working with highly dilute solutions.
• Polyamines stimulate the activity of RNase I.
• RNase I is less stable against heat inactivation than RNase A.

Related Products

Proteinase K RNase A/T1 Mix GeneJET™ Plasmid Miniprep Kit Bovine Serum Albumin Water, nuclease-free

References

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

Updated kovo 18, 2008 09:57