Klenow DNA Polymerase is a truncated version of E. coli DNA polymerase I, which lacks the 5'->3' exonuclease activity. Since it contains intact 3'->5' proofreading exonuclease activity in addition to 5'->3' polymerase, its fidelity of DNA synthesis equals to that of the full-length E. coli DNA polymerase I. It also contains strand displacement activity for nick translation.
- Molecular weight: 68 KDa
- Reaction temperature: 25℃
- Heat inactivation: 75℃, 20 min
- Specific activity: 20,000 units/mg
- 5’→3’ exonuclease: No
- 3’→5’ exonuclease: Yes
- Strand Displacement: Yes
- Radioisotope labeling of double stranded DNA with recessed 3' ends.
- Filling-in of 5' overhangs to produce blunted duplex ends
- Removal of 3' overhangs to produce blunted duplex ends
- Second strand synthesis of cDNA obtained by reverse transcriptase
- Radioisotope labeling of DNA with random primers
- Purity: >99% on SDS-PAGE
One unit is defined as the amount of enzyme required to incorporate 10 nmol of dNTP into acid-insoluble materials with 70 mg/ml of denatured herring sperm DNA as template in 30 min at 37℃.
100 mM KPO4 (pH 6.5), 1 mM DTT, 50% glycerol
10X Klenow DNA Polymerase buffer
100 mM Tris-HCl (pH 7.9), 100 mM MgCl2, 10 mM DTT, 500 mM NaCl
- 10X Klenow DNA Polymerase buffer
- Sterile water