AcuGenix™ AmpHifi HS DNA Polymerase I

High-Fidelity Hot-Start Polymerase for Specific, Robust PCR Workflows

~71× fidelity

Compared with wild-type Taq

Up to 10 kb

From human genomic DNA

> 80% GC

Local GC content tolerated

AcuGenix™ AmpHifi HS DNA Polymerase I, BR3P101 product image

Product overview

AcuGenix™ AmpHifi HS DNA Polymerase I is a thermostable, high-fidelity DNA polymerase engineered via directed evolution and protein engineering. It delivers exceptional amplification specificity and fidelity, making it an ideal choice for sensitive molecular biology applications requiring minimal sequence bias.

Features and applications

Features

Ultra-High Fidelity with Proofreading Activity: Possesses robust 3′→5′ exonuclease proofreading activity that corrects nucleotide misincorporation in real time, resulting in an extremely low mismatch rate during PCR.
Antibody-Blocked Hot-Start Specificity: Modified with a thermolabile monoclonal antibody that fully inhibits 5′→3′ polymerase and 3′→5′ exonuclease activity at room temperature. This prevents non-specific amplification and primer-dimer formation prior to the initial denaturation step, drastically improving assay specificity.
Optimized Buffer with Extension Cofactor: The proprietary reaction buffer incorporates a dedicated extension cofactor that maintains stable, high-efficiency amplification across diverse template types, simplifying protocol optimization for routine experiments.
User-Friendly Workflow: Enables reliable performance for both novice and expert users, with minimal optimization required for standard cloning, long-fragment amplification, and other common PCR applications.

Applications

Routine gene cloning and sub-cloning
Long-fragment PCR for complex genomic templates
NGS library preparation (amplicon and shotgun libraries)
Site-directed mutagenesis and SNP genotyping
Amplification of GC-rich or secondary structure-laden templates
Research-use diagnostic assay development

Components

Component	Specification/Notes
5×HiFi Buffer I	5×
AmpHifi HS DNA Polymerase I	1 U/ μL

Note: 5×HiFi Buffer I is the base buffer. Consult sales technical staff for optimized buffers.

Performance data

Fidelity Assay

BR3P101 AcuGenix™ AmpHifi HS DNA Polymerase I: Figure 1. Superior fidelity of AmpHifi HS DNA Polymerase I — **Figure 1. Superior fidelity of AmpHifi HS DNA Polymerase I.**

Libraries were amplified with AmpHifi HS DNA Polymerase I, Supplier K, or Supplier Y polymerases after standardized library construction. Normalized sequencing data revealed lower SNP counts for AmpHifi HS, confirming higher fidelity. AmpHifi HS DNA Polymerase I demonstrated ~71-fold higher fidelity than wild-type Taq polymerase, making it suitable for applications requiring minimal amplification errors.

Processivity

BR3P101 AcuGenix™ AmpHifi HS DNA Polymerase I: Figure 2. Robust long-range synthesis by BR3P101 across 1–10 kb targets — **Figure 2. Robust long-range synthesis by BR3P101 (AmpHifi HS DNA Polymerase I).** Human genomic DNA (gDNA) was amplified with BR3P101 and the optimized buffer across 1–10 kb targets. The system consistently yielded specific amplicons up to 10 kb in length, demonstrating high processivity and suitability for long-fragment amplification workflows.

High-GC Tolerance

BR3P101 AcuGenix™ AmpHifi HS DNA Polymerase I: Figure 3. Robust amplification of high-GC targets — **Figure 3. Robust amplification of high-GC targets.**

Human gDNA was amplified using AmpHifi HS DNA Polymerase I and the optimized buffer. Lanes 1–3 show products from target regions with 74%, 73%, and 72% GC content. The system efficiently amplified targets with local GC content >80% and overall GC content >70%, confirming its suitability for challenging GC-rich templates in NGS and PCR applications.