Thymine-DNA Glycosylase (TDG)

Unit Definition

One unit is defined as the amount of enzyme required to create an AP site from 1 pmol of a 60-mer oligonucleotide duplex containing a single T:G mispair within a CpG context* in a total reaction of 20 µl in 1 hour at 37°C.

*A T:G mispair is prepared by having the T located at the 24th position of a 5′ FAM-labelled 60 mer ssDNA and a
G at the 25^th position of an unlabeled complement, annealed to form a duplex substrate.

Reaction Conditions

1X TDG Reaction Buffer
Incubate at 37°C

1X TDG Reaction Buffer
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
0.25 mM TCEP
10 µg/ml Recombinant Albumin
(pH 7.9 @ 25°C)

Storage Buffer

10 mM Tris-HCl
500 mM NaCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation