The PR1001-HF-500 is the 500-reaction larger pack of the Accuris High Fidelity 2X Master Mix — a pre-formulated single-tube 2× concentrated PCR master mix that combines the High Fidelity polymerase, buffer with PCR enhancers, dNTPs, and MgCl₂ in one tube. The HF master-mix format is the right pick for production cloning, sequencing template prep, and mutagenesis workflows where the convenience of pre-formulated chemistry matters and the workflow runs at moderate-to-mid throughput. Ships as 5 × 1.25 mL tubes (500 reactions at 25 µL master mix per 50 µL reaction).
Key Specifications
| SKU | PR1001-HF-500 |
| Type | High Fidelity 2X Master Mix — pre-formulated single-tube HF PCR master mix |
| Reactions per pack | 500 reactions (pack format: 5 × 1.25 mL tubes) |
| Concentration | 2× master mix (25 µL master mix + 25 µL primers/template/water = 50 µL standard reaction) |
| Polymerase | High Fidelity DNA Polymerase (low-error-rate proofreading enzyme) |
| Buffer composition | PCR enhancers + dNTPs + MgCl₂ pre-formulated for HF chemistry |
| PCR product format | Blunt-end (HF polymerases lack the terminal-A overhang activity of standard Taq) |
| Smaller pack sister | PR1001-HF-200 (200 reactions, 2 × 1.25 mL) |
| HF + Hot Start sister | PR1001-HFHS-500 (HF + Hot Start combined master mix) |
| Standalone HF enzyme sister | PR1000-HF-200 / PR1000-HF-1000 (HF Polymerase, separate format) |
| Standard Taq Master Mix sister | PR1001-200 / PR1001-1000 (lower fidelity, lower cost) |
| Returns | Non-returnable — perishable reagent per Accuris reagent-return policy |
HF Master Mix vs Standalone HF Enzyme — Workflow Decision
| Format | Workflow setup | When to pick |
| PR1001-HF-500 (this product) — HF Master Mix | Combine 25 µL master mix + 25 µL (primers / template / water) → ready to cycle | Routine HF cloning workflow; production sequencing-prep; mutagenesis at moderate throughput; standardized HF buffer across reactions |
| PR1000-HF-1000 — HF standalone enzyme | Combine separate HF enzyme + 5X buffer (with dNTPs) + primers + template → ready to cycle | Custom-buffer workflows; specialty MgCl₂ adjustments; cost-controlled enzyme volume |
The master-mix format is the workflow-fastest pick — fewer pipetting steps, standardized chemistry across reactions, reduced operator error. Standalone enzyme is the right pick when the workflow needs custom MgCl₂ optimization for difficult templates or when bulk enzyme procurement at lower per-unit cost is the priority.
Why HF Polymerase Master Mix
Standard Taq DNA Polymerase has a measurable error rate during DNA synthesis. For workflows where individual PCR product molecules will be cloned, sequenced, or used as functional templates, the polymerase error rate matters because errors accumulate into the cloned, sequenced, or functional output. HF polymerases reduce error rate by 10–100× vs standard Taq through 3'→5' exonuclease proofreading activity that detects and corrects misincorporation errors during synthesis.
The HF Master Mix delivers HF chemistry at the same workflow ergonomics as standard Taq Master Mix — same setup steps, same number of pipetting operations, same final reaction volume.
Blunt-End PCR Products — Cloning Implication
HF polymerase produces blunt-end PCR products suitable for blunt-end cloning vectors. Standard Taq adds a 3'-terminal A overhang for TA-cloning compatibility; HF polymerase lacks this activity because the proofreading exonuclease removes terminal mismatches.
Workflow implication: blunt-end cloning vectors accept direct ligation of HF PCR products; TA-cloning vectors require a post-PCR A-tailing step (separate Taq or A-tailing reagent) before TA ligation; and restriction-site cloning proceeds by restriction-digesting the PCR product first, unaffected by the HF choice.
HF vs HF + Hot Start — Variant Decision
The Accuris HF master-mix family ships in two variants:
| Variant | Hot Start? | When to pick |
| PR1001-HF (this product's family) | No | Routine HF workflows on well-optimized templates; reactions assembled on ice |
| PR1001-HFHS | Yes | Challenging templates needing both HF and Hot Start specificity (low-copy GC-rich cloning, multiplex HF, room-temperature setup workflows) |
For routine cloning / sequencing-prep where the template is well-characterized and reactions are assembled on ice, the PR1001-HF master mix is the right pick. For demanding templates (GC-rich, low-copy, multiplex) that benefit from both HF and Hot Start, step up to PR1001-HFHS.
PR1001-HF Pack-Size Family
| SKU | Reactions | Pack format | Use case |
| PR1001-HF-200 | 200 | 2 × 1.25 mL | Smaller pack — pilot HF work, occasional cloning |
| PR1001-HF-500 (this product) | 500 | 5 × 1.25 mL | Production HF workflows — cloning core, sequencing-prep, mutagenesis production |
The 5-tube format of the 500-reaction pack supports rotating tubes through active use while keeping the rest in deep storage.
Reagent Composition
Per the Accuris Life Science Reagents Catalog, each PR1001-HF-500 pack includes the High Fidelity 2X Master Mix pre-formulated with:
- High Fidelity DNA Polymerase
- Buffer with PCR enhancers
- dNTPs
- MgCl₂
Standard reaction setup: combine 25 µL of HF master mix + 25 µL of (primers + template + water) in a 50 µL reaction. The pre-formulated chemistry is optimized for HF reaction kinetics — typical extension time per cycle is longer than standard Taq (HF polymerases are slower-but-more-accurate enzymes).
Application Profile
Per the Accuris Life Science Reagents Catalog, HF polymerase applications span blunt-end cloning (direct ligation into blunt-end vectors), TA cloning (with a post-PCR A-tailing step for TA-vector compatibility), colony PCR where fidelity matters, fast PCR, GC- and AT-rich DNA amplification on challenging-composition templates where proofreading helps, and low-copy PCR on low-input templates where accurate amplification matters. The brochure highlights HF polymerase as "Ideal for cloning (blunt end), mutagenesis, & microarray applications."
When to Pick This Pack
Order PR1001-HF-500 when:
- Production cloning bench — cloning core facilities or biotech cloning teams running HF PCR daily for ongoing cloning projects, where master-mix convenience matters
- Sequencing template prep at production scale — facilities running HF PCR for Sanger or NGS amplicon-based sequencing template prep
- Mutagenesis production workflow — site-directed mutagenesis benches where every reaction requires HF
- Antibody-engineering / protein-engineering workflows — workflows generating cloned variants of expression constructs at production scale
- Standardized HF chemistry across multiple operators — multi-operator labs where master-mix standardization reduces protocol variability
Ordering & Configuration
Storage & handling. Per Accuris kit instructions — typical HF master-mix storage at −20 °C with limited freeze-thaw cycle tolerance. The 5-tube format supports rotating tubes through active use.
Cloning-vector compatibility. Blunt-end products work directly with blunt-end vectors; TA-cloning workflows need post-PCR A-tailing.
Hot Start consideration. For challenging templates (GC-rich, low-copy, multiplex), evaluate stepping up to PR1001-HFHS-500 which combines HF + Hot Start chemistry.
Returns. Per the universe price-list category header: reagent products including PCR master mixes are non-returnable due to perishable nature.
Confirm at quote: downstream workflow (cloning / sequencing / mutagenesis), pack-size match (200 vs 500 reaction), HF vs HF+HS variant decision, cloning-vector format, shipping conditions (cold-chain), and lead time.
Warranty: Per Accuris Life Science Reagents standard kit warranty terms — confirm at quote.
Accuris PR1001-HF High Fidelity Master Mix family:
- PR1001-HF-200 — 200 reactions, 2 × 1.25 mL
- PR1001-HF-500 (this product) — 500 reactions, 5 × 1.25 mL (production-throughput)
- PR1001-HFHS-200 / PR1001-HFHS-500 — HF + Hot Start combined master mix family
Pick HF master mix for routine cloning / sequencing-prep / mutagenesis workflows. Step up to HF+HS for challenging templates needing both proofreading and hot-start specificity. Step out to standalone PR1000-HF for custom-buffer workflows.
