DNA Polymerase I

#3003

 

300 units

#3003L

 

1,500 units

DESCRIPTION
DNA Polymerase I is DNA- dependent DNA polymerase that consists of a single polypeptide chain (Mr = 109,000). This enzyme can function as a 5'3' DNA polymerase, a 5'3' exonuclease, and a 3'5' exonuclease.

SOURCE
Recombinant E. coli strain.

APPLICATIONS
Labeling of DNA by nick translation.
Second strand cDNA synthesis.
End-labeling of DNA molecules with protruding 3' tails.

UNIT DEFINITION
One unit is defined as the amount of enzyme required to catalyze the incorporation of 10nmol of dNTPs into acid-insoluble form in 30 minutes at 37.

STANDARD UNIT ASSAY CONDITIONS
67mM KPO4 (pH7.5), 6.7mM MgCl2, 1mM DTT, 133M activated calf thymus DNA and 33M dATP, dCTP, dGTP and [3H]-dTTP.

CONCENTRATION :5-10 units/l.

10X REACTION BUFFER
100mM Tris-HCl (pH7.5@37), 50mM MgCl2, 7.5mM DTT. dNTPs not included.

STORAGE CONDITIONS
50mM Tris-HCl (pH7.5), 1mM DTT, 0.1mM EDTA and 50% glycerol. Store at -20.

REFERENCES
1. Meinkoth, J. and Wahl, G.M., (1987) Methods Enzymol. 152, 91.
2. Kelley, W.S. and Stump, K.H. (1979) J. Biol. Chem. 254, 3206. 3. Richardson, C.C. et al. (1964) J. Biol. Chem. 239, 222.