Human DNA Polymerase Gamma

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Description

• The simplest DNA polymerase known in both size and catalysis (1)

Applications

• Mitochondrial enzyme used for testing the toxicity of drugs on critical human oxidative functions and testing replication of mitochondrial DNA

Unit Definition

One unit is the amount of enzyme required to incorporate 1 nmole of total nucleotide into acid-insoluble form in 60 minutes at 37°C

Reaction Buffer

50 mM Tris-HCl (pH 8.7)
10 mM MgCl₂
0.4 mg/ml bovine serum albumin
100 mM KCl
15% glycerol

Storage Buffer

20 mM Tris-HCl (pH 8.0)
1.0 mM dithiothrietol
0.1 mM EDTA
0.2 M NaCl
50% (v/v) glycerol

Assay Conditions

50 mM Tris-HCl (pH 8.7)
10 mM MgCl₂
0.4 mg/ml bovine serum albumin
1 mM dithiothreitol
100 mM KCl
15% glycerol
0.05 mM of each dCTP, dGTP, dTTP, and [α-³²P]dATP
10 μg activated DNA
Incubation for 15 minutes at 37°C in a total reaction volume of 50 μl

Quality Control

All preparations are assayed for contaminating endonuclease and exonuclease activities

Storage Conditions

Store at -20°C
Product shipped on dry ice

References

(1) Abbotts, J., SenGupta, D.N., Zmudzka, B., Widen, S.G., Notario, V., and Wilson, S.H. (1988) Biochemistry 27, No. 3, 901-909