Ultra-Pure Phi29 DNA Polymerase
| Cat. No. | E014 | ||||||
| Name | Ultra-Pure Phi29 DNA Polymerase | ||||||
| Unit | 100 µl | ||||||
| Category | Molecular Biology Enzymes and Kits | ||||||
| Description |
Ultra-Pure Phi29 DNA Polymerase is a highly processive enzyme known for its powerful strand-displacement activity, making it ideal for isothermal amplification of both circular and linear DNA templates. It is particularly effective in techniques such as Rolling Circle Amplification (RCA), Multiple Displacement Amplification (MDA), and Whole Genome Amplification (WGA). Thanks to its 3'→5' exonuclease activity, Ultra-Pure Phi29 DNA Polymerase boasts exceptional fidelity, enabling the amplification of even very small quantities of DNA templates. The enzyme undergoes an advanced, multi-step purification process that utilizes a combination of physical, chemical, and enzymatic methods to ensure the complete removal of contaminating genomic DNA. Rigorous quality control is performed, including a non-specific DNase Activity Assay and a qPCR DNA Contamination Test using a TaqMan probe, ensuring superior purity and performance.
1Buffer contains 0.5 μg/μl BSA. Keep at -20°C at all times except when using the product. |
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| Application |
• Rolling circle amplification (RCA) |
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| Format General | Enzyme supplied with 5X Reaction Buffer | ||||||
| Storage Buffer | 50 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT, 100 mM NaCl, and 50% (v/v) Glycerol. | ||||||
| Storage Condition | Store all components at -20°C. | ||||||
| Caution | This product is distributed for laboratory research only. Not for diagnostic use. | ||||||
| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. E014 |
| How can I design gene-specific primer (to replace the random hexamer) to use with the Phi29? | |
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There are no specific requirements for designing gene specific primers for use with Phi29 amplification. As Phi29 functions at a low temperature, the Tm and length is not a problem.
Please be aware that you should use more primer in the reaction than with regular PCR. The final concentration of the primers should be 50uM.
In addition, you will see better results if the primers are 3' end protected.
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| What types of DNA templates can be amplified with this polymerase? | |
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This enzyme efficiently amplifies circular and linear DNA templates, making it ideal for applications like Rolling Circle Amplification (RCA), Multiple Displacement Amplification (MDA), and Whole Genome Amplification (WGA).
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| How long should the reaction be incubated? | |
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The reaction should be incubated at 30°C for 4 to 16 hours. The optimal time depends on the template length and the desired amplification yield.
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| What precautions should be taken to prevent contamination? | |
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To minimize contamination:
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| How do I inactivate the enzyme after the reaction? | |
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The enzyme can be inactivated by incubating the reaction at 65°C for 10 minutes, followed by cooling to 4°C.
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| What is the source of this enzyme? | |
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The enzyme is produced recombinantly in E. coli, which has been engineered to express the enzyme gene. While the original gene may come from another organism, all production and purification occur using E. coli under controlled conditions.
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