Engineered DNA Polymerases

Ramon Kranaster
myPOLS Biotec UG (haftungsbeschränkt), Universitätsstr. 10, 78457 Konstanz, Germany,

By combinatorial protein design myPOLS Biotec is evolving new generations of DNA polymerases for advanced applications. As an example, we recently developed HiDi DNA polymerase that is an engineered DNA polymerase developed for approaches in which high discrimination between matched and mismatched primers is required (e.g., HLA analysis, allele-specific PCR (ASA) or methylation-specific PCR (MSP)). Whereas many DNA polymerases tolerate mismatched primers at least to some extent, HiDi DNA polymerase efficiently discriminates those and specifically produces amplicons in case of perfectly matched primer pairs only. This renders HiDi DNA polymerase the prime enzyme for SNP detections, HLA genotyping or the analysis of single CpG methylation sites in MSP. Based on HiDi DNA polymerase, we provide custom-made advanced and reliable qPCR mixes. Furthermore, since HiDi DNA polymerase allows allele-specific PCRs directly from oral swabs without requiring DNA isolation or any other tedious sample processing step, a Direct SwabPCR 2x Master Mix was established.
We also developed Volcano DNA polymerase – an engineered, extremely thermostable enzyme that has both, reverse transcriptase and PCR activity. Volcano DNA polymerase has a half-life at 95°C of >40 min and it facilitates “zero-step” RT-PCRs directly from RNA templates (without an isothermal reverse transcription step). These properties allow reverse transcription reactions at high temperatures, thus minimizing the problems encountered with strong secondary structures in RNA that only melt at elevated temperatures.
myPOLS Biotec is actively pursuing research on DNA polymerases to shape their properties for advanced applications. In the next months new DNA polymerases will be introduced by myPOLS Biotec that include:
• DNA polymerases resistant to known inhibitors (e.g., from blood or plants) allowing direct PCR without tedious sample preparations
• DNA polymerases enabling multiplex DNA and RNA diagnostics
• DNA polymerases that are tolerant to high salt conditions
• DNA polymerases allowing PCR from damaged DNA samples
• DNA polymerases that tolerate highly modified nucleotides
• Temperature sensitive DNA polymerases with intrinsic hot start function
Still missing the right DNA polymerase for your applications? We will assist you in establishing your approach using our wide knowledge on DNA polymerase engineering. With our expertise we tailor DNA polymerases for your needs! Contact us!

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