| Cat# | POL-005 |
| MW | 68 kDa (Reducing) |
| Label | His Tag |
| Notes | The active reducing agent in the reaction buffer is critical for this enzyme. Although the reaction buffer supplied with the enzyme contains DTT, in order to ensure maximum activity, 4 mM DTT should be added when using buffers that have been stored for long periods of time or buffers that have been repeatedly freeze-thawed. Reaction temperatures above 65°C are not recommended. The enzyme does not have 5′→3′ nucleic acid exonuclease activity. |
| Purity | ＞95% by SDS-PAGE and HPLC |
| Source | Bacillus subtilis phage phi29 (Φ29) |
| Storage | Store at -25 ~ -15°C for 2 years. |
| Activity | 10 U/μL |
| Synonyms | phi29 DNA Polymerase, DNA polymerase |
| Component | 10 U/μL phi29 DNA Polymerase, 100 mM KCl, 10 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Tween 20, 50% Glycerol, 0.5% NP40, pH 7.4 at 25°C |
| Description | phi29 DNA polymerase is a DNA polymerase cloned from Bacillus subtilis phage phi29 (Φ29). |
| Applications | This enzyme has excellent strand replacement and sustained synthesis capabilities, enabling the unstranding and replication of complex DNA structures and isothermal DNA polymerization reactions in vitro that do not depend on thermal cycling. This enzyme possesses 3′→5′ nucleic acid exonuclease proofreading activity. |
| Specification | 250 U; 1250 U |
| Dilution Buffer | 100 mM KCl, 10 mM Tris-HCl, 0.1 mM EDTA, 1 mM DTT, 0.5% Tween 20, 50% Glycerol, 0.5% NP40, pH 7.4 at 25°C |
| Expression System | E.coli |
| 10× Reaction Buffer | 50 mM Tris-HCl, 10 mM MgCl₂, 10 mM (NH₄)₂SO₄, 4 mM DTT, (pH 7.5 at 25°C). |
| Activity Definitions | One unit refers to the amount of enzyme required to catalyze the incorporation of 0.5 pmol of dNTP into an acid-insoluble material in 10 minutes at 30°C. |
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