DNA Polymerase I Large (Klenow -exo) Fragment is a DNA-dependent DNA polymerase that lacks both the 5' to 3´ and 3' to 5’ exonuclease activity of intact E. coli DNA Polymerase I. DNA Polymerase I Large (Klenow -exo) Fragment is used for random priming and chain displacement amplification. But it is not recommended in blunt formation for blunt-ended ligation.
DNA Polymerase I Large (Klenow -exo) Fragment is purified from a recombinant E.coli strain.
Concentration & Storage Condition
- 5 unit/㎕. Store at -20℃.
- 50 mM Tris-HCl, pH 7.5, 1 mM DTT, 0.1 mM EDTA, 50% (v/v) glycerol.
10x Reaction Buffer
- 500 mM Tris-HCl, pH 7.2, 100 mM MgSO₄, 1 mM DTT.
- One unit is defined as the amount of enzyme that incorporates 10 nmole of total dNTP into TCA-insoluble material in 30 min at 37°C.
The reaction conditions are: 67 mM potassium phosphate, pH 7.5, 6.7 mM MgCl₂, 1 mM DTT, 50 ㎍/ml activated calf thymus DNA and 33 μM each of dATP, dCTP, dGTP and dTTP (a mix of unlabeled and [³H]dTTP).
- Activity, exo and endonuclease activity test, SDS-PAGE purity, performance tests.