Size: 200 Units
Klenow Fragment is a proteolytic product of intact DNA polymerase I which retains
5'-3' DNA polymerase and 3'-5' exonuclease activity but lacks 5'-3' exonuclease
Recombinant E. coli strain.
- DNA sequencing by the Sanger dideoxy method.
- Filling-in and/or labeling of recessed 3'-ends.
- Second strand cDNA synthesis.
- End-labeling of DNA molecules with protruding 3' tails.
- Synthesis of dsDNA from ssDNA templates during in vitro mutagenesis.
Unit (U) 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°C.
Standard Unit Assay Conditions:
67mM KPO4 (pH7.5), 6.7mM MgCl2, 1mM DTT, 133µM activated calf thymus
DNA and 33µM dATP, dCTP, dGTP and [3H]-dTTP.
10X Reaction Buffer:
100mM Tris-HCl (pH7.5@37°C), 50mM MgCl2, 7.5mM DTT. dNTPs not included
50mM Tris-HCl (pH7.5), 1mM DTT, 0.1mM EDTA and 50% glycerol. Store at -20°C.
Klenow Fragment (5U/ul)