MagniPhi® DNA Polymerase | 500025

Phi29 DNA polymerase is an extremely processive DNA polymerase (up to more than 70 kb per binding event) with extraordinary strand displacement capacity, exhibiting 3’ → 5’ proofreading exonuclease activity, resulting in an exceptionally high synthesis fidelity. These distinctive features make this DNA polymerase the perfect choice for isothermal DNA amplification or any application requiring a high fidelity DNA polymerase with strong strand displacement capacity, such as Rolling Circle Amplification (RCA).

Applications

• Highly accurate DNA synthesis
• Rolling Circle Amplification (RCA)
• Whole Genome Amplification (WGA)
• Amplification of DNA from filter paper blood spot samples
• Protein-primed DNA amplification
• RNA-primed DNA amplification
• In situ genotyping with padlock probes
• Recombination based cloning
• Proximity ligation assay

Figure 1: 4BB™ MagniPhi^® benchmarking against leading market competitors in whole genome amplification

Quality Control

Absence of endonucleases and exonucleases:

4BB™ MagniPhi^® DNA polymerase has been determined to be free of detectable endonucleases, exonucleases and nicking activity. A fluorogenic substrate designed to react with all these kinds of nucleases has been incubated in the presence of 1μg of polymerase enzyme for one hour at 37°C. No increase in fluorescence compared with the negative control was detected.

Absence of ribonucleases:

4BB™ MagniPhi^® DNA polymerase has been determined to be free of detectable single-strand ribonuclease activity. A fluorogenic substrate designed to react with these kinds of nucleases has been incubated in the presence of 1μg of the enzyme for 30 min at 37°C. No increase in fluorescence compared with the negative control was detected.

Purity:

The purity of the enzyme has been determined to be higher than 95% by SDS polyacrylamide gel electrophoresis and densitometric measurements.

Functional assay

4BB™ MagniPhi^® DNA polymerase has been tested for exponential amplification of 10pg and 1ng of high molecular weight human genomic DNA. In the absence of DNA input (non-template control, NTC) no background amplification was exhibited under similar conditions.

Shipping and Storage

4BB™ MagniPhi^® DNA  polymerase is shipped on dry ice. On receipt, the kit should be stored immediately at -20^oC in a non-frost-free (constant temperature) freezer. When stored correctly, the product can be kept for a minimum of six months after shipping without displaying any reduction in performance. For longer periods, store the kit at -80^oC. Avoid repeated freeze-thaw cycles.