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
| SKU | PR1001-HFHS-200 |
| Type | Single-tube 2X HF + Hot Start combined master mix |
| Pack quantity | 200 reactions (2 × 1.25 mL tubes) |
| PCR mode | High Fidelity + Hot Start (combined chemistry) |
| Master mix composition | HF polymerase (with proofreading) + antibody-mediated hot-start chemistry + buffer + PCR enhancers + dNTPs + MgCl₂ |
| Sister SKUs | PR1001-HFHS-500, PR1001-HFHS-S |
When to Pick HFHS vs HF-Only vs Hot Start-Only
The PR1001 master mix family includes three related chemistry variants:
| SKU family | Chemistry | When to pick |
| PR1001-HF-200 / -500 | HF only (no hot-start) | Workflows needing HF accuracy without hot-start activation control (cloning, sequencing-prep, mutagenesis at well-optimized protocols) |
| PR1001-HS-200 / -1000 | Hot Start only (standard Taq + hot-start) | Workflows needing hot-start activation control without HF accuracy (multiplex, GC-rich, low-copy at standard accuracy) |
| PR1001-HFHS-200 (this product) | HF + Hot Start combined | Workflows needing both — multiplex HF cloning, GC-rich HF amplification, low-copy HF where pre-activation suppression matters |
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.
