AcuGenix™ AmpHifi HS DNA Polymerase III

Next-Generation Hot-Start High-Fidelity Polymerase for Demanding PCR & NGS Workflows

~150× fidelity

Compared with wild-type Taq

1–20 kb

Processive long-fragment synthesis

Up to 80% GC

Down to pg-level inputs

Product overview

AcuGenix™ AmpHifi HS DNA Polymerase III is a next-generation hot-start high-fidelity DNA polymerase engineered via site-directed mutagenesis and domain recombination. It delivers significantly enhanced DNA binding affinity and processivity, alongside exceptional amplification compatibility with GC-rich templates and trace template inputs. Validated by standardized mismatch rate assays, its replication fidelity is approximately 150-fold higher than that of wild-type Taq DNA polymerase, making it an ideal choice for high-accuracy molecular biology applications.

Features and applications

Features

Next-Generation Engineered Performance: Optimized via targeted protein engineering to outperform conventional high-fidelity polymerases in specificity, yield, and template tolerance.
Superior DNA Binding & Processivity: Enhanced affinity for template DNA enables sustained synthesis of long fragments per binding event, reducing amplification bias for complex templates.
Challenging Template Compatibility: Robust performance with high-GC targets (up to 80% local GC) and trace input amounts (down to pg-level templates), minimizing the need for protocol optimization.
Ultra-High Fidelity: ~150× higher replication fidelity than wild-type Taq polymerase, drastically reducing sequence errors for downstream sequencing, cloning, and diagnostic applications.
Streamlined Workflow: Hot-start design prevents non-specific amplification at room temperature, enabling room-temperature reaction assembly with minimal optimization.

Applications

NGS library preparation (amplicon, shotgun, and target enrichment libraries)
Low-input/low-template PCR for single-cell, cfDNA, or FFPE samples
Amplification of GC-rich, repetitive, or secondary structure-laden templates
High-fidelity gene cloning and site-directed mutagenesis
SNP genotyping and variant detection assays
Research-use diagnostic assay development

Components

Components	Specification/Notes
5×HiFi Buffer III	5×
AmpHifi HS DNA Polymerase III	1 U/ μL

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

Performance data

Fidelity Assay

BR3P103 AcuGenix™ AmpHifi HS DNA Polymerase III: Figure 1. Fidelity Benchmarking Against Competitor Enzymes — **Figure 1. Fidelity Benchmarking Against Competitor Enzymes.** BR3P103 (AmpHifi HS DNA Polymerase III) exhibits a lower mutation frequency than Supplier K, Supplier V, and Supplier Y. Its amplification fidelity is roughly 150-fold higher than that of wild-type Taq DNA polymerase, demonstrating outstanding high-fidelity performance.

Note: BR3P103 is the catalog number for this product. Lower mutation frequency directly correlates with reduced base misincorporation during amplification, minimizing false positives in downstream applications including NGS variant calling, gene cloning, and diagnostic assay development. This makes the enzyme ideally suited for workflows requiring minimal amplification errors.

Processivity Assay

BR3P103 AcuGenix™ AmpHifi HS DNA Polymerase III: Figure 2. Amplification of Targets Across Varying Lengths — **Figure 2. Amplification of Targets Across Varying Lengths.** When paired with the optimized amplification buffer, BR3P103 supports processive synthesis of 1–20 kb target amplicons, making it well-suited for long-fragment PCR workflows.

GC-rich Template Compatibility Assay

BR3P103 AcuGenix™ AmpHifi HS DNA Polymerase III: Figure 3. Compatibility with GC-Rich Templates — **Figure 3. Compatibility with GC-Rich Templates.** Lanes 1–5 correspond to multiplex target amplicons with GC content ranging from 30%–80%. Amplification assays were performed on templates with varying GC content using BR3P103 and its optimized buffer, demonstrating efficient amplification across a broad GC content range.

Note: BR3P103 is a specific catalog number for this polymerase, corresponding to a different pack size of the same enzyme as BR3P101 validated for long-fragment amplification, with identical performance characteristics. Low misincorporation rates minimize false positives in NGS variant calling, gene cloning, and diagnostic assay development.