DNA Polymerase I, Klenow Fragment from Escherichia coli
SIGMA/D8276 - buffered aqueous glycerol solution
CAS Number: 9012-90-2
MDL Number: MFCD00283091
Product Type: Chemical
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| concentration | ~3,000 units/mL |
| foreign activity | Endonuclease, none detected |
| form | buffered aqueous glycerol solution |
| grade | Molecular Biology |
| mol wt | 103 kDa |
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | −20°C |
| UniProt accession no. | P00582  |
| Analysis Note: | The activity is assayed in a reaction mixture containing 50 mM potassium phosphate (pH 7.5), 3 mM MgCl2, 1 mM 2-mercaptoethanol, 32.5 μM 32P-dATP, 32.5 μM dTTP, 62.5 μg/ml poly(dA-dT) and 0.01-1 unit enzyme. |
| Application: | Suitable for: • DNA sequencing by the Sanger dideoxy method • Synthesis of the complementary strand of cDNA • Filling in 5′-overhangs in double stranded DNA to form blunt ends • Mutagenesis of DNA with second strand synthesis using oligonucleotides • Labeling DNA by the random primer method |
| General description: | DNA polymerase I yields two fragments (small and large) upon protease digestion. The large fragment (Klenow fragment) loses the 5′ exonuclease activity that is present in the intact holoenzyme. However, it retains both the polymerase 5′→3′ activity and the 3′→5′ exonuclease activity of the native enzyme. |
| Other Notes: | DNA Polymerase I is supplied as a solution in 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 5 mM dithiothretol, and 50% glycerol (v/v) . |
| Other Notes: | One unit converts 10 nanomoles of deoxyribonucleoside triphosphates into acid insoluble material in 30 min. at 37 °C. |
| Preparation Note: | The enzyme solution may be diluted with 50 mM Tris-HCl, pH 7.5, 100 mM ammonium sulfate, 10 mM 2-mercaptoethanol, and 1 mg/ml bovine serum albumin. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 2 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352204 |