| Description |
Klenow Fragment is a truncated verson of E coli DNA Polymerase I. The enzyme exhibits 5'→3' polymerase (DNA synthesis) and proofreading (3′→5′) nuclease activities, but lacks 5'→3' exonuclease activity. |
| Applications |
- Removeal of 3´ overhangs to form blunt ends
- Random-primed DNA labeling
- Labeling by fill-in 5'-overhangs of dsDNA to form blunt ends
- DNA sequencing by the Sanger method
- Site-specific mutagenesis of DNA with synthetic oligonucleotides
- Second strand synthesis of cDNA
|
| Source |
A recombinant E. coli strain carrying the Klenow Fragment of E coli PolA gene. |
| Unit Definition |
1 unit is defined as the amount of polymerase required to convert 10 nmol of dNTPs into acid insoluble material in 30 minutes at 37°C. |
| Components |
- Klenow Fragment: 5,000 U/ml in 25 mM Tris-HCl, 1 mM DTT, 0.1 mM EDTA, 50% Glycerol, pH 7.4 @ 25°C
- 10X Reaction Buffer: 500 mM NaCl, 100 mM Tris-HCl, 100 mM MgCl2, 10 mM DTT, pH 7.9 @ 25°C
|
| Quality Control |
- The absence of endodeoxyribonucleases confirmed by appropriate quality test.
- Functionally tested for fill in of 5'-overhanging DNA termini and for random primed DNA labeling.
|
| Storage Condition |
-20 °C |