NEBuilder HiFi DNA Assembly enables virtually error-free joining of DNA fragments, even those with 5´- and 3´-end mismatches. This flexible kit 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.
4 Reasons to choose NEBuilder HiFi
- Save time – Enjoy simple and fast seamless cloning in as little as 15 minutes.
- Flexibility – Use one system for both "standard-size" cloning and larger gene assembly products, up to 6 fragments.
- Compatible with downstream applications – DNA can be used immediately for transformation or as template for PCR or RCA.
- Adaptable – Adapts easily for multiple DNA manipulations, including site-directed mutagenesis.
NEBuilder HiFi DNA assembly offers unique features over other DNA assembly methods, including:
- 3´- and 5´-end mismatch removal
- Bridging of two ds-fragments with a ssDNA oligo
- Improved methods for site-directed mutagenesis
How does NEBuilder HiFi stack up?
Performance data and advantages against NEB Gibson Assembly
Performance data and advantages against In-Fusion HD
Synthetic Biology/DNA Assembly Selection Chart
Comparison of DNA Assembly Reaction Types
Not your average DNA assembly reagent
NEBuilder HiFi DNA Assembly can be used for a variety of DNA assembly methods.
NEBuilder HiFi productsNEBuilder HiFi DNA Assembly Cloning Kit (NEB #E5520)
- 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
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 email@example.com. The use of these products may require you to obtain additional third party intellectual property rights for certain applications.
GIBSON ASSEMBLY® is a registered trademark of Synthetic Genomics, Inc.
IN-FUSION® is a registered trademark of ClonTech Laboratories, Inc.
Protocols for NEBuilder® HiFi DNA Assembly
- NEBuilder HiFi DNA Assembly Reaction Protocol
- NEBuilder® HiFi DNA Assembly Chemical Transformation Protocol
- NEBuilder® HiFi DNA Assembly Chemical Transformation Protocol (E2621)
- NEBuilder® HiFi DNA Assembly Electrocompetent Transformation Protocol
- NEBuilder® HiFi Electrocompetent Transformation Protocol (E2621)
- Construction of an sgRNA Cas9 expression vector
- Improved method for assembly of linear yeast expression cassette using NEBuilder® HiFi DNA Assembly Master Mix
- Improved methods for site-directed mutagenesis using NEBuilder® HiFi DNA Assembly Master Mix
- Synthetic Biology/DNA Assembly Selection Chart
Other Tools & Resources
While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at firstname.lastname@example.org.
This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Did you know that one of the advantages of NEBuilder HiFi is that it removes 3’ and 5’ -end mismatch sequences prior to fragment assembly? Learn how!
Learn how you can use NEBuilder HiFi to generate an sgRNA-Cas9 expression vector with a single-stranded oligo bridge.
Find out how NEBuilder® HiFi DNA Assembly can reliably join DNA fragments in a single tube, isothermal reaction, with advantages over NEB Gibson Assembly®.
Primer Design and Fragment Assembly Using NEBuilder® HiFi DNA Assembly or Gibson Assembly® E5510 E5520
Watch an interactive tutorial on primer design to see how simple it really is to clone with either NEBuilder HiFi DNA Assembly or the Gibson Assembly Cloning Kit.
Learn how NEBuilder® HiFi DNA Assembly bridges dsDNA with a ssDNA oligo.