A wild type form of the high fidelity DNA polymerase Phi29 DNA polymerase
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 -20oC 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 -80oC. Avoid repeated freeze-thaw cycles.
