RNase III

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