Klenow Fragment, 3'->5' exo–, is the Large Fragment of E.coli DNA Polymerase I with a molecular weight of 68kDa. It exhibits 5'->3' polymerase activity, but lacks the 3'->5' exonuclease activities. The 3'->5' exonuclease activity of the enzyme is eliminated by mutations in the 3'->5' exonuclease active site.
- Random priming labeling
- DNA sequencing by the Sanger dideoxy method
- Second strand cDNA synthesis
- Second strand synthesis in mutagenesis protocols
20 mM Tris-HCl
100 mM NaCl
1.0 mM DTT
0.1 mM EDTA
pH 7.5 @ 25°C
Supplied with: 10x Blue Buffer
10x Blue Buffer:
pH 7.9 @ 25 ºC
1 unit is defined as the amount of enzyme that will incorporate 10 nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30min at 37°C.
Recommended Storage Condition: -20°C