Endonuclease III (Nth)

Unit Definition

One unit is defined as the amount of enzyme required to cleave 1 pmol of a 34 mer oligonucleotide duplex containing a single AP site* in a total reaction volume of 10 µl in 1 hour at 37°C in 1X Endonuclease III Reaction Buffer containing 10 pmol of fluorescently labeled oligonucleotide duplex.

* An AP site is created by treating 10 pmol of a 34 mer oligonucleotide duplex containing a single uracil residue with 1 unit of Uracil-DNA Glycosylase (UDG) for 2 minutes at 37°C.

Reaction Conditions

1X Endonuclease III (Nth) Reaction Buffer
Incubate at 37°C

1X Endonuclease III (Nth) Reaction Buffer
20 mM Tris-HCl
1 mM EDTA
1 mM DTT
(pH 8 @ 25°C)

Usage Concentration

10,000 units/ml

Storage Temperature

-20°C

Storage Conditions

10 mM Tris-HCl
250 mM NaCl
1 mM DTT
0.1 mM EDTA
200 μg/ml BSA
50% Glycerol
pH 7.4

Heat Inactivation

65°C for 20 min

Unit Assay Conditions

1X Endonuclease III (Nth) Reaction Buffer containing 10 pmol of fluorescently labeled oligonucleotide duplexes in a total reaction volume of 10 µl