DNA Polymerase I, E. coli

Catalog No. 1080

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Price From: $60.00

Quick Overview

Mesophilic DNA-dependent DNA polymerase with inherent 3'→5' and 5'→3' exonuclease activity

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Price From: $60.00

Details

Source

Escherichia coli

Description

  • Exhibits the 5'→3' polymerase activity
  • Exhibits the 5'→3' exonuclease activity, active only on duplex DNA
  • Contains the 3'→5' exonuclease, primarily active on single-stranded DNA (1)
  • Ultrapure recombinant enzyme

Applications

  • Used to prepare radioactive probes by nick translation (2) and random priming (3)
  • Useful for end-labeling of DNA molecules wih 3' and 5' protruding tails or blunt-ended

Unit Definition

One unit is the amount of enzyme required to incorporate 10 nmoles of total deoxyribonucleotide into acid-insoluble material in 30 minutes at 37°C with DNase I-activated DNA as the template primer

Storage Buffer

50 mM potassium phosphate (pH 7.0)
0.25 mM dithiothreitol
50% (v/v) glycerol

Assay Conditions

67 mM potassium phosphate (pH 7.4)
6.7 mM MgCl2
1 mM dithiothreitol
0.033 mM of each dCTP, dGTP, dTTP and [α-32P] dATP
4.5 µg activated DNA
Incubation is for 30 minutes at 37°C in a total reaction volume of 100 µl

Quality Control 

All preparations are assayed for contaminating endonuclease activity. Typical preparations are greater than 95% pure, as judged by SDS polyacrylamide gel electrophoresis

Storage Conditions

Store at -20ºC
Product shipped on dry ice 

Downloads

Certificate of Analysis PDF -Current Lot
MSDS PDF     

References

1. Lehman, I.R., Enzymes 14, 15-37. 1981
2. Rigby, P.W.J., Diekmann, M., Rhodes, C. and Berg, P., J. Mol. Biol. 113, 237-251. 1977
3. Hartman, C.P. and Robussay, D., Gene Amplification and Analysis (Chirikjian, J.G. and Pappas, T.S., eds.) 2, 17-39, Elsevier/North Holland, New York. 1981