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F1-X™Next Generation Gibson Assembly 1-Step Master Mix
You deserve more value from your DNA assembly reagent. Try F1-X risk-free.
Cost-Saving
15% savings over competitors*
NGS-Verified
Whole-Plasmid Sequencing on Every Batch
Benchmarked
Head-to-Head vs. NEBuilder®, GeneArt™ & More
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100% Money-Back Guarantee
*Competitor pricing sourced from manufacturer website as of March 2026, subject to change.
F1-X™ DATA
Performance Highlights
Every claim on this page is backed by whole-plasmid sequencing data — not just colony counting. We’re getting the whole story (not just counting colonies, or sequencing the insert).
Experimental benchmarks across multiple assembly conditions.
A — Head to Head Data
Benchmarked against leading competitors for error free clone rate
4-fragment assembly · 40 bp overlaps · whole-plasmid NGS sequencing · n=2 biological replicates · 7 colonies per replicate
F1-X is optimized specifically for cloning fidelity (high correct-clone rates) in multi-fragment assemblies. F1-X may produce >3x fewer colonies than competitor kits, especially before optimization of transformation conditions. Here, we measure what matters: getting an abundance of colonies to screen, and the percentage of colonies that are correct. DNA assembly and sequencing error rates are construct dependent.
HOW WE COMPARE
Head-to-Head Comparison
F1-X™ brings you multi-fragment power at a fraction of the cost.
| Metric | F1-X™ | NEBuilder® HiFi | GeneArt™ Gibson Assembly® HiFi (Invitrogen) | Gibson Assembly® Cloning Kit (NEB) |
|---|---|---|---|---|
| Number of fragments | ✓2–12 (up to 14 demonstrated)¹ | 2–6 recommended (up to 11 demonstrated) | Up to 5 + vector (≤6 total) | 2–6 recommended (up to 12 demonstrated) |
| Cloning fidelity (internal testing)² | ✓1.8× | 1.5× | 1.3× | 1.0× (baseline) |
| Pricing (per reaction, 10-rxn kit) | ✓$15.80/rxn | $18.60/rxn | $16.40/rxn | $19.40/rxn |
| Reaction time | 15–60 min | 15–60 min | 15–60 min | 15–60 min |
| Reaction temperature | ✓50°C (50–56°C range) | 50°C (40–50°C range) | 50°C | 50°C (40–50°C range) |
| Reaction volume | ✓20 µL standard (verified to 2.5 µL) | 20 µL | 20 µL | 20 µL |
| Max total assembly size | ✓Up to 100 kb | ~19 kb documented | Not specified | ~20 kb documented |
| Positive control type | ✓4-fragment GFP (visual readout) | 2-fragment dsDNA | 2-fragment Kan/Amp selection | 2-fragment dsDNA |
| Temperature optimization range | ✓50–56°C | 40–50°C | 50°C only | 40–50°C |
| Restriction enzyme seam removal | ✓Yes — eliminates RE site overhangs during assembly³ | Yes — removes 3′/5′ end flaps including RE seams | Not specified | Not specified |
| Crude PCR tolerance | Up to 20% v/v | Up to 20% v/v | Direct from PCR supported | Up to 20% v/v |
← scroll for full comparison →
1 Up to 14 fragments demonstrated in a lycopene biosynthesis pathway assembly. See: F1-X™ Application Note: Multi-Fragment Assembly of a Lycopene Biosynthesis Pathway (Racer Biosciences, 2026).
2 Cloning fidelity expressed as a ratio of NGS-verified error-free clone rate relative to the lowest-performing product (Gibson Assembly® Cloning Kit = 1.0× baseline). Racer Biosciences internal testing: n=2 biological replicates, 4-fragment assembly, 40 bp overlaps. Seven colonies per replicate analyzed by whole-plasmid next-generation sequencing. DNA assembly and sequencing error rates are construct dependent.
3 F1-X™ eliminates restriction enzyme site overhangs (e.g., EcoRI seams) during assembly via exonuclease chew-back and seamless ligation. See F1-X™ User Guide, Appendix B.
Visual GFP Positive Control
F1-X™ includes a 4-fragment GFP positive control with a visual green fluorescent readout of both assembly and transformation success. Competitors use simple 2-fragment controls — a more rigorous QC tool that directly demonstrates multi-fragment assembly capability.
Extended Temperature Range (50–56°C)
F1-X™ operates across 50–56°C, enabling higher-stringency annealing for GC-rich or complex overlap sequences. NEBuilder HiFi operates at 40–50°C and GeneArt specifies 50°C only — providing an optimization lever that reduces mis-priming in complex assemblies.
Demonstrated 14-Fragment Assembly
F1-X™ supports 2–12 fragments per reaction, with up to 14 fragments demonstrated in a lycopene biosynthesis pathway assembly. NEBuilder HiFi documents up to 11 inserts but recommends ≤5. GeneArt caps at 5 fragments plus a vector.
Up to 100 kb Assembly Size
F1-X™ documentation specifies total assembly sizes up to 100 kb. NEBuilder HiFi has documented ~19 kb total. Gibson Assembly has documented ~20 kb total. A significant differentiator for synthetic biology users building large pathways or genetic circuits.
TECHNICAL DETAILS
Guidelines
Reaction Conditions
Temperature
50°C (50–56°C range)
Time (2–3 Fragments)
15 min
Time (4–12 Fragments)
60 min
Reaction Volume
20 µL standard (2.5–20 µL range)
DNA Input
1–10 fmol/µL
Approx. ng/µL per fragment (20 µL rxn): 0.5 kb: 1–2 ng/µL · 1 kb: 0.5–1.25 ng/µL · 10 kb: 1.25–2.5 ng/µL
Assembly Range
Fragments
2–12 per reaction
Fragment Size
100 bp – 32 kb per fragment
Total Assembly Size
Up to 100 kb
Overlap (2–3 Frags)
20–40 bp
Overlap (4+ Frags)
40+ bp
Stability & Compatibility
Storage
−20°C
Shelf Life
12 months at −20°C (unopened)
Crude PCR Compatible
Yes (≤20% v/v)
Automation Compatible
Yes
WORKFLOW
Three Steps
From fragments to verified constructs with no special equipment.
Combine Fragments
Add fragments to F1-X™ Master Mix in a single tube.
Incubate
50°C for 15–60 min.
Transform
Transform directly into competent cells or use for cell-free workflows.
WHAT'S INCLUDED
Every Kit Contains
Each F1-X kit includes the master mix and control components sized to the selected reaction count.
F1XGA10R
F1-X™ Next-Generation 1-Step Gibson Assembly® Master Mix — 10 reactions
F1XGA50R
F1-X™ Next-Generation 1-Step Gibson Assembly® Master Mix — 50 reactions
F1XGA100R
F1-X™ Next-Generation 1-Step Gibson Assembly® Master Mix — 100 reactions
F1XGA250R
F1-X™ Next-Generation 1-Step Gibson Assembly® Master Mix — 250 reactions
| SKU | Product | Component SKU | Volume | Price / Action |
|---|---|---|---|---|
| F1XGA10R | F1-X™ Next-Generation 1-Step Gibson Assembly® Master Mix — 10 reactions | $158 | ||
| F1-X™ Master Mix (2×) | F1XMM10R | 1 × 100 µL | ||
| F1-X™ Positive Control (2×) | F1XCTRL | 1 × 50 µL | ||
| F1XGA50R | F1-X™ Next-Generation 1-Step Gibson Assembly® Master Mix — 50 reactions | $625 | ||
| F1-X™ Master Mix (2×) | F1XMM50R | 1 × 500 µL | ||
| F1-X™ Positive Control (2×) | F1XCTRL | 1 × 50 µL | ||
| F1XGA100R | F1-X™ Next-Generation 1-Step Gibson Assembly® Master Mix — 100 reactions | $1,173 | ||
| F1-X™ Master Mix (2×) | F1XMM100R | 1 × 1,000 µL | ||
| F1-X™ Positive Control (2×) | F1XCTRL | 1 × 50 µL | ||
| F1XGA250R | F1-X™ Next-Generation 1-Step Gibson Assembly® Master Mix — 250 reactions | $2,358 | ||
| F1-X™ Master Mix (2×) | F1XMM250R | 1 × 2,500 µL | ||
| F1-X™ Positive Control (2×) | F1XCTRL | 1 × 50 µL | ||
Resources & Downloads
F1-X delivers excellent results in multi-fragment cloning projects. I get numerous, sequence-correct clones. The workflow is straight-forward and easy to use. I also appreciate the lower cost per reaction compared with other suppliers that I have been using for years.
Kent Boles
CEO, Bespoke Bioworks, Inc.
USE CASES
Where F1-X™ Excels
If you need to assemble DNA, F1-X can help.
Click a category to explore F1-X use cases.
Synthetic Biology & Pathway Engineering
Strain Engineering
Metabolic pathway prototyping and biosynthetic cluster assembly require reliable multi-fragment joins. Racer assembly tools reduce junction errors and preserve design logic across complex constructs.
Synthetic Biology & Gene Circuits
Multi-part genetic circuits, toggle switches, and biosensors depend on scarless, high-fidelity assembly. F1-X™ maintains design integrity across complex multi-fragment constructs.
Large Genomic Constructs
BAC-scale constructs and large genomic inserts require reliable joins across 5–10+ kb fragments. Racer assembly tools are recommended up to 100 kb.
For research use only. Not for use in diagnostic procedures.
F1-X™ technology is protected under US patent numbers 7,776,532, 8,435,736, 8,968,999, and additional patents pending.
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