DNA Polymerase I, Large (Klenow) Fragment
| Catalog # | Pack size | Price, Euro | SHOP |
| 401002 | 200 u | 40,00 | Add to cart |
| 401010 | 1000 u | 120,00 | Add to cart |
Description:
DNA Polymerase I, Large (Klenow) Fragment is a proteolytic product of <em>E. coli</em> DNA Polymerase I which retains polymerization and 3´-->5´ exonuclease activity, but has lost 5´-->3´ exonuclease activity. Klenow retains the polymerization fidelity of the holoenzyme without degrading 5´ termini.
Source:
<em>E.coli</em> strain harboring the plasmid that directs the synthesis of Klenow fragment
Reaction Buffer:
(1X) 10 mM Tris-HCl (pH 7.5), 5 mM MgCl2, 5 mM dithiothreitol
Klenow Fragment is also active in any restriction enzyme reaction buffer and T4 DNA Ligase reaction buffer when supplemented with dNTPs.
Quality Assurance:
Purified free of contaminating endonucleases and exonucleases.
Unit Definition:
One unit is defined as the amount of enzyme required to convert 10 nmols of dNTPs to an acid-insoluble form in 30 minutes at 37°C.
Concentration:
5,000 - 50,000 units/ml.
Storage Conditions:
100 mM KPO4 (pH 6.5), 1 mM dithiothreitol and 50% glycerol. Store at -20°C.
Applications:
- DNA sequencing by the Sanger dideoxy method
- Fill-in of 5´ overhangs to form blunt ends
- Removal of 3´ overhangs to form blunt ends
- Second strand cDNA synthesis
- Second strand synthesis in mutagenesis protocols
