NEBuilder^® HiFi DNA Assembly

Assembling DNA fragments is a key part of both synthetic biology techniques and cloning. NEBuilder HiFi DNA Assembly enables virtually error-free joining of DNA fragments, even those with 5´- and 3´-end mismatches. This flexible mix enables simple and fast seamless cloning utilizing a proprietary high-fidelity polymerase. Find out why NEBuilder HiFi is the next generation of DNA assembly and cloning.

5 Reasons to choose NEBuilder HiFi 

1. Save time – Enjoy simple and fast seamless cloning in as little as 15 minutes.
2. Flexibility – Use one system for both "standard-size" cloning and larger gene assembly products, up to 12 fragments.
3. Compatible with downstream applications – DNA can be used immediately for transformation or as template for PCR or RCA.
4. Adaptable – Adapts easily for multiple DNA manipulations, including site-directed mutagenesis. 
5. Save money - No licensing fees from NEB for NEBuilder products

Want to try NEBuilder HiFi DNA Assembly?

 

Request a Sample 

NEBuilder HiFi DNA assembly offers unique features over other DNA assembly methods, including:

• Remove 3´- or 5´-end restriction digest sites or other mismatches during HiFi assembly 
• Bridge two ds-fragments with a ssDNA oligo
• Perform multi site-directed mutagenesis
• Assemble multiple annealed ssDNA oligos

 
How does NEBuilder HiFi stack up?

Performance data and advantages against GeneArt^® Gibson Assembly^® and In-Fusion^® Snap Assembly 
Synthetic Biology/DNA Assembly Selection Chart
Comparison of DNA Assembly Reaction Types
Guidelines for using NEBuilder HiFi DNA Assembly
High-throughput cloning and automation solutions

 
Not your average DNA assembly reagent

NEBuilder HiFi DNA Assembly can be used for a variety of DNA assembly methods.

 
NEBuilder HiFi products

• Includes NEB 5-alpha competent cells
• NEB recommends NEB 5-alpha Competent E. coli (High Efficiency, NEB #C2987) for routine assemblies of 15 kb or less
• For repetitive sequences, NEB recommends NEB Stable Competent E. coli (NEB #C3040)

• Includes NEB 10-beta competent cells
• NEB recommends NEB 10-beta Competent E. coli (High Efficiency, NEB #C3019) or NEB 10-beta Electrocompetent E. coli (NEB #C3020) for assemblies larger than 15 kb

• Does not include competent cells 

Online Supporting Tools

	NEBuilder® Assembly Tool Use to easily design primers for your NEBuilder HiFi DNA Assembly reactions.
	NEBaseChanger®  For multi-site mutagenesis primer design using NEBuilder HiFi DNA Assembly.

One or more of these products are covered by patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc. For more information, please email us at busdev@neb.com. The use of these products may require you to obtain additional third party intellectual property rights for certain applications. 

Application Notes

• Improved methods for site-directed mutagenesis using NEBuilder HiFi DNA Assembly Master Mix
• Improved method for assembly of linear yeast expression cassettes using NEBuilder HiFi DNA Assembly Master Mix
• Bridging dsDNA with a ssDNA Oligo and NEBuilder HiFi DNA Assembly to create an sgRNA-Cas9 Expression Vector
• Nanoliter Scale DNA Assembly Utilizing the NEBuilder HiFi Cloning Kit with the Labcyte^® Echo^® 525 Liquid Handler
• Modular DNA Assembly of PIK3CA Using Acoustic Liquid Transfer in Nanoliter Volumes
• Miniaturized Multi-Piece DNA Assembly Using the Echo 525 Liquid Handler