Accuris High Fidelity Hot Start Master Mix, 200 Reactions

The PR1001-HFHS-200 is the single-tube 2X High Fidelity + Hot Start combined master mix, sold as 200-reaction pack (2 × 1.25 mL tubes) per the Accuris Life Science Reagents Catalog. The HFHS master mix combines two PCR-chemistry advantages in one reagent: (a) High Fidelity proofreading polymerase for low-error-rate amplification (~1×10⁻⁶ vs standard Taq's ~1×10⁻⁴) AND (b) antibody-mediated hot-start activation for pre-activation suppression in sensitive workflows (multiplex, GC-rich, low-copy).

Key Specifications

When to Pick HFHS vs HF-Only vs Hot Start-Only

The PR1001 master mix family includes three related chemistry variants:

Pick HFHS when:

• The workflow needs HF accuracy AND multiplex / GC-rich / low-copy hot-start support
• Sample → cloning workflows with primer-pair multiplexing where sequence accuracy AND activation control both matter
• Method-development PCR where reducing both error sources (chemistry + activation non-specifics) matters

Pick HF-only when: the workflow is HF-grade but hot-start premium is not justified (well-optimized single-primer-pair PCR). Pick Hot Start-only when: the workflow needs activation control but standard Taq accuracy is sufficient.

Per Brochure HFHS Applications

The PR1001-HFHS family combines the application profiles of the HF and Hot Start individual chemistries. From HF chemistry (proofreading) it covers blunt-end / TA cloning, sequencing-prep PCR with a low error rate, site-directed mutagenesis for sequence-accuracy preservation, and GC- and AT-rich amplification. From Hot Start chemistry (antibody-mediated) it covers multiplex PCR with non-specific suppression, low-copy PCR with no pre-activation extension, and primer-dimer-prone setups. The combined HFHS applications are workflows where multiple constraints apply at once: multiplex HF cloning, GC-rich HF mutagenesis, and low-copy HF sequencing-prep.

HFHS vs Standalone HF + Hot Start Manual Combination

Some labs attempt to combine standalone HF polymerase with separate Hot Start activation reagents. The PR1001-HFHS combined master mix offers significant advantages: the antibody-mediated hot-start is engineered to match the HF polymerase chemistry (ad-hoc combinations may have suboptimal activation kinetics), it delivers single-tube convenience with no multi-reagent setup, its combined chemistry is quality-controlled as a unit by Accuris rather than operator-assembled, and it reduces workflow complexity through fewer pipetting steps and procurement lines.

Pick the engineered HFHS master mix over manual HF + Hot Start combination for any workflow where the combined chemistry is needed.

Application Profile

Multiplex HF PCR for cloning. Multiplex amplification of multiple targets with downstream cloning where both multiplex specificity (Hot Start) and cloning fidelity (HF) matter.

GC-rich amplification with downstream sequencing. GC-rich templates that need both Hot Start specificity (template secondary structure handling) and HF accuracy (sequencing-grade output).

Low-copy HF amplification. Sensitive PCR from low template where both Hot Start sensitivity and HF accuracy across cycles matter.

Sequencing-prep multiplex. NGS library prep multiplexing where both primer-pair non-specific suppression and library-sequence accuracy matter.

Method-development PCR optimization. Protocol-development workflows where reducing both chemistry error and activation non-specifics matters for protocol robustness.

Pharma / biotech screening with downstream sequencing. Drug-discovery PCR pipelines where sequence accuracy of amplified targets matters downstream.

Ordering & Configuration

Per-reaction workflow consumable. Each PCR reaction uses 12.5 µL of the 2X HFHS master mix (confirm exact volume in product insert). A 1.25 mL tube supports 100 reactions; the 200-reaction pack is 2 × 1.25 mL tubes.

Storage. HFHS reagent storage applies. Confirm storage protocol at order.

Pair with template / primers / water. Operator supplies template, primers, water; the master mix has everything else.

Confirm at quote: workflow combined HF+Hot Start chemistry need, throughput (drives 200-rxn vs 500-rxn pack-size selection), evaluation interest, comparison to HF-only or Hot Start-only chemistries, and lead time.

Warranty: Per Accuris standard reagent terms. Confirm at quote.

Accuris PR1001-HFHS combined HF + Hot Start master mix family:

- PR1001-HFHS-S (Sample, 5 reactions) first-use evaluation

- PR1001-HFHS-200 (this product): 200 reactions

- PR1001-HFHS-500: 500 reactions

Pick the HFHS master mix when the workflow needs both HF accuracy and Hot Start activation control; sister chemistries (HF-only PR1001-HF, Hot Start-only PR1001-HS) cover single-chemistry needs.