| Source | Isolated from E.coli strain that carries the cloned DNA polymerase I (klenew fragment) gene |
| Description | DNA Polymerase I, Large (Klenow) Fragment is a proteolytic product of E. coli DNA Polymerase I which retains polymerization and 3’ -> 5’ exonuclease activity, but has lost 5’ -> 3’ exonuclease activity.
Applications:
- dideoxy sequencing;
- polishing ends;
- second strand cDNA synthesis
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| Unit | One unit is the amount of enzyme required to incorporate 10 nmol of dNTPs into an acid-insoluble material in 30 minutes at 37°C.
Unit Assay Conditions: 1 x Klenow Buffer, 33 µM dNTP including [3H]-dTTP and 70 µg/ml denatured calf thymus DNA.
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| Reaction buffer | SE-buffer Klenow fragment, (50 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 5 mM DTT.) |
| Optimal temperature | 37oC |
| Storage conditions | 10 mM KH2PO4 (pH 7.5); 50 mM KCl; 0.5 mM EDTA; 7 mM 2-mercaptoethanol; 50% glycerol. Store at -20°C. |
| Quality control | Purified free of contaminating endonucleases and exonucleases. |
| MSDS: | Download MSDS as PDF |
