RNase III

Catalog No. 1340

Quick Overview

Divalent metal-dependent nuclease that cleaves long double-stranded RNA (dsRNA) into short dsRNAs (13-30 bases)

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Price From: $105.00

Details

Source

E. coli plasmid with mc gene of E. coli RNase III

Description

  • RNase III is a divalent metal dependent nuclease that cleaves long double-stranded RNA (dsRNA) into short dsRNAs (13-30 bases). 
  • Products of RNase III mimic siRNA structures produced by Dicer enzyme (5'-PO4, 3'-OH and a dinucleotide 3' overhang). This unique feature enables to generate a population of RNAs that, after transfection into mammalian cells, can induce RNAi (1-4).

Applications

  • Digestion of long dsRNA to short dsRNA
  • Used in RNA structure studies (5)
  • Transfection of RNase III cleavage products can be used to induce RNAi in mammalian cells
  • Used in RNA processing and maturation studies (6-8)

Unit Definition

One unit of RNase III is the amount of enzyme required to digest 1 µg of dsRNA to siRNA in 20 minutes at 37°C in a total reaction volume of 50 µl.

Concentration

2 units/µl

Reaction Buffer

300 mM Tris-HCl (pH 8.0 at 25°C)
1.6 M NaCl
10 mM dithiothreitol
1 mM EDTA 

Storage Buffer

30 mM Tris-HCl (pH 8.0 at 25°C)
500 mM NaCl
1 mM dithiothreitol
0.5 mM EDTA
50% (v/v) glycerol

Quality Control

All preparations are assayed for contaminating exonuclease, endonuclease, for nonspecific RNase and single- and double-stranded DNase activities. Typical preparations are greater than 90% pure, as judged by SDS polyacrylamide gel electrophoresis.

Storage Conditions

Store at -20°C
Shipped on dry ice

References

1. Yang, D. et al. (2002) Proc. Natl. Acad. Sci. USA, 99, 9942-9947.
2. Calegari, F. et al. (2002) Proc. Natl. Acad. Sci. USA, 99, 14236-14240.
3. Donze, O. and Picard, D. (2002) Nucleic Acids Res, 30, e46. Laboratory, Cold
    Spring Harbour.
4. Morlighem, J.E. et al. (2007) Biotechniques, 42, 599-606.
5. Evguenieva-Hackenberg, E. and Klug, G. (2000) J. Bacteriol. 182, 4719.
6. Nicholson, A. (1999) FEMS Microbiol. Rev. 23, 371.
7. Drider, D. et al. (1999) J. Mol. Microbiol. Biotechnol. 1, 337.
8. Grunberg-Manago, M. (1999) Annual Rev. Genet. 33, 193.