Inside high-fidelity PCR: accuracy, applications, and innovation

  • Published: 15 May 2025
We are proud to present Solis Hot Start HiFi DNA Polymerase, a high-performance, high-fidelity enzyme designed for precision-demanding PCR applications such as sequencing, cloning, and mutagenesis. This product has superior performance with a variety of amplicons, can be used for long-range and challenging PCR, and has broad multiplexing capabilities.

What is a high-fidelity (HiFi) DNA polymerase?
High-fidelity polymerases are specialized DNA polymerase enzymes designed to replicate DNA with exceptionally low error rates. They possess strong 3' to 5' exonuclease (proofreading) activity, allowing them to detect and correct mismatched nucleotides during DNA synthesis. A common source for HiFi polymerases is Pyrococcus furiosus (Pfu), known for its thermostability and superior proofreading capacity.

Where are HiFi DNA polymerases used?
Using these types of polymerases is important in research and diagnostics, where minimizing mutations is critical. One rapidly growing field is sequencing, both Next Generation Sequencing (NGS) and Sanger sequencing. HiFi polymerases are used there for either library preparation or sequencing platforms themselves.

Other application possibilities include accurate cloning for protein expression, PCR amplification, CRISPR-Cas9 and site-directed mutagenesis.

What is special about Solis Hot Start HiFi DNA Polymerase?
Solis Hot Start HiFi DNA Polymerase is a novel in silico designed polymerase with 3′→5′ proofreading activity demonstrating enhanced fidelity of more than 100 times compared to wild-type Taq, as determined by state-of-the-art single-molecule real-time (SMRT) sequencing. Additionally, our experts have engineered it to efficiently incorporate U nucleotides. That is critical in work with damaged DNA and bisulfite sequencing. Of course, it can be used in all other sequencing workflows that require high-fidelity polymerases as well.

Solis Hot Start HiFi DNA Polymerase is highly accurate, convenient to use and robust thanks to proprietary Stability TAG technology. It can also amplify challenging templates with up to 79% GC content, it is capable of highly multiplex reactions and the enhanced processivity of the polymerase allows for long-range PCR.

In addition, a strong chemical hot start has been implemented for the polymerase to improve specificity while minimizing mispriming and extension from non-specific annealed primers and primer-dimers, simplifying highly multiplexed reactions.

The bottom line
Whether you are working on library preparation for your sequencing experiment and require highly accurate PCR for cloning, protein expression or site-directed mutagenesis. Whether you are studying DNA methylation or are faced with a challenging template, make sure to try out Solis Hot Start HiFi DNA Polymerase and test the great features in your application!

For more details, watch our WEBINAR.