Large Fragment of DNA Polymerase I
recombinant, E. coli
For general laboratory use.
Unit Definition: One unit is defined as the amount of enzyme required to convert 10 nmoles of dNTPs to an acid insoluble form in 30 minutes at 37 °C.
Shipping: shipped on gel packs
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles
Shelf Life: 12 months
Form: liquid (Supplied in 100 mM KPO4 pH 6.5, 1 mM DTT and 50 % [v/v] glycerol)
Concentration: 5 units/μl
Klenow Fragment is the large fragment of DNA Polymerase I that retains its 5'→3' polymerase, 3'→5' exonuclease and strand displacement activities. The enzyme lacks the 5'→3' exonuclease activity of intact DNA polymerase I. Klenow retains the polymerization fidelity of the holoenzyme without degrading 5' termini.
Incubate for 15 min. at 25 °C
Stop reaction by alternativly
10x Reaction Buffer:
500 mM Tris-HCl pH 7.6 at 25 °C, 50 mM MgCl2 and 10 mM DTT.
Excessive amounts of enzyme or longer reaction times may result in recessed ends due to the 3'→5' exonuclease activity of the enzyme.
The enzyme is greater than 98 % pure as indicated by SDS-polyacrylamide gel electrophoresis and contains no detected endonuclease activity. Incubation of 10 units of Klenow with supercoiled plasmid DNA produced no nicked molecules after 20 hours at 37 °C as determined by agarose gel electrophoresis analysis.
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