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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'¡æ3'
DNA polymerase, a 5'¡æ3' exonuclease, and a 3'¡æ5' exonuclease.
SOURCE
Recombinant E. coli strain.
APPLICATIONS
¡Ü Labeling of DNA by nick
translation.
¡Ü Second strand cDNA synthesis.
¡Ü 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¡É.
STANDARD UNIT ASSAY CONDITIONS
67mM
KPO4 (pH7.5), 6.7mM MgCl2, 1mM DTT, 133¥ìM activated calf thymus DNA and
33¥ìM dATP, dCTP, dGTP and [3H]-dTTP.
CONCENTRATION :5-10 units/¥ìl.
10X REACTION BUFFER
100mM Tris-HCl
(pH7.5@37¡É), 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¡É.
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. |