Phi29 DNA Polymerase
|E014||1000 U (100 μl)|
|Description||Phi29 DNA Polymerase is a highly processive polymerase which exhibits a strong strand-displacement function. These functions allow for highly efficient isothermal amplification of circular or linear DNA templates via rolling circle amplification (RCA), multiple displacement amplification (MDA) and/or whole genome amplification (WGA). Phi29 DNA Polymerase has extremely high fidelity due to its inherent 3’→5’ exonuclease activity and can amplify from very small amounts of starting templates.|
|Unit quantity||1000 U (100 μl)|
• Rolling circle amplification (RCA) • Multiple displacement amplification (MDA) • Whole genome amplification (WGA) • DNA template preparation for sequencing • Protein-primed DNA amplification
|Format||Enzyme supplied with 5X Reaction Buffer|
Store all components at -20°C.
|Storage Buffer||50 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT, 100 mM NaCl, 0.5% Tween-20, 0.5% NP-40 and 50% (v/v) Glycerol.|
One unit is defined as the amount of Phi29 DNA Polymerase that is required to incorporate 0.5 pmol of dNTP into acid insoluble material in 10 minutes at 30°C.
|How can I design gene-specific primer (to replace the random hexamer) to use with the Phi29?|
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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