Product Information

RNase III, E.coli is a homodimeric, divalent metal ion dependent nuclease. RNase III cleaves long double-stranded RNA (dsRNA) into short 12-30 base dsRNAs containing a 5'-PO₄, 3'-OH, and a dinucleotide 3' overhang.

Source: Recombinant E.coli
Concentration: 0.5U/μL
Unit Definition: One unit is the amount of enzyme required to digest 1μg of dsRNA to siRNA in 20 minutes at 37℃ in a total reaction volume of 50μL.
Storage Buffer: 500mM NaCl, 20mM Tris-HCl (pH 8.0, 25℃), 0.5mM EDTA, 0.5mM DTT, 50% Glyercol.

Companion Products:

Cat#ON-078

10X RNaseIII Reaction Buffer

inside with:

- 500mM Tris-HCl (pH7.5, 25℃)
- 500mM NaCl
- 10mM DTT

Key points of operation

Recommended protocol:

In ice, combine the following, in order:

Component

Amount

ddH₂O

X μL

10X RNase III Reaction Buffer

10μL

dsRNA

X μL (10μg)

RNase III

10-20 units

10X MnCl₂

10μL

Total Volume

100μL

Mix and incubate for 20 minutes at 37℃.
Add 10μL 10X EDTA to stop the reaction.

Security Information

Storage Conditions:
Long term (Infrequent use): -70℃
Daily/Weekly use: -20℃*
Do not store in a frost-free freezer.

RNase III stores at -20℃ may result in the formation of haze. If this happens, keep the tube at room temperature until tube temperature is up to 4~10℃. Then mix gently to clarify the solution. Haze should disappear. And there is negligible loss of activity.

Quality Assurance: Free of Endonuclease, Exonuclease, and Nickase activities. Pass functional testing
Physical Purity: ＞90% by SDS-PAGE.

Reference

1. Hugh D. Robertson, Robent E. Webster, and Norton D. Zinder. The Journal of Biological Chemistry. 243, 82-91(1968).
2. Neelam Srivastava and Rai Ajit K Srivastava. Biochemistry and Molecular Biology International. 39, 171-180(1996).

Enzyme & Related Buffer for mRNA

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