DNA Polymerase I Large (Klenow) Fragment is the large fragment of E. coli DNA Polymerase I. The Klenow Fragment retains the DNA-dependent DNA polymerase activity of the E. coli DNA Polymerase I but lacks the 5’→3’ exonuclease activity. With its inherent 3’→5’ exonuclease activity, Klenow possesses the polymerization fidelity of the holoenzyme without degrading 5'-termini.
Applications:
- DNA blunting by filling-in 5'-overhangs with unlabeled or labeled dNTPs
- cDNA second-strand synthesis
- Generate single-stranded DNA probes using random primers
- Site-directed DNA mutagenesis using synthetic oligonucleotides
- Dideoxy DNA sequencing of single- or double-stranded DNA templates
- 3’→5’ exonuclease activity can blunt a 3’-overhang
Concentration: 5 U/μl
Storage: Store all components at -20°C.
Storage Buffer: 25 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT, and 50% (v/v) Glycerol.
Note:
- One unit is defined as the amount of DNA Polymerase I Large (Klenow) Fragment that catalyzes the incorporation of 10 nmol of dNTP into acid insoluble material in 30 minutes at 37°C using poly(dA-dT):poly(dA-dT) as a template:primer.
- This product is distributed for laboratory research only. Caution: Not for diagnostic use.
