Home Applications Cloning & Synthetic Biology DNA Assembly and Cloning NEBuilder® HiFi DNA Assembly

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 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.

5 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 11 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.
Save money - No licensing fees from NEB for NEBuilder products

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

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, useful for both synthetic biology and cloning.

NEBuilder_Overview_Infographic_0916

NEBuilder HiFi products

NEBuilder 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)

NEBuilder HiFi DNA Assembly Bundle for Large Fragments (NEB #E2623)

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

NEBuilder HiFi DNA Assembly Master Mix (NEB #E2621)

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 busdev@neb.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.

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FAQs for NEBuilder® HiFi DNA Assembly

How does overlap length affect NEBuilder HiFi DNA Assembly efficiency?

Protocols for NEBuilder® HiFi DNA Assembly

Application Notes for NEBuilder® HiFi DNA Assembly

Legal Information

This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

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 gbd@neb.com.

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.

Videos

NEBUILDER HIFI DNA ASSEMBLY^® Removal of 3´ end Mismatches

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!
Construction of an sgRNA-Cas9 Expression Vector via an ssOligo Bridge

Learn how you can use NEBuilder HiFi to generate an sgRNA-Cas9 expression vector with a single-stranded oligo bridge.
NEBUILDER^® HiFi: the next generation of DNA Assembly

In this webinar, we will explore some of the latest applications for NEBuilder HiFi DNA Assembly. View full Q & A summary.
Introduction to NEBUILDER HIFI DNA ASSEMBLY^®

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^®

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.
NEBUILDER^® HIFI DNA ASSEMBLY^® : Bridging dsDNA with a ssDNA Oligo

Learn how NEBuilder® HiFi DNA Assembly bridges dsDNA with a ssDNA oligo.
NEBUILDER^® Assembly Tool 2.0 What’s New?

NEBuilder Assembly Tool can be used to design primers for your NEBuilder HiFi or Gibson Assembly reactions based on entered fragment sequences and the polymerase being used for amplification. This video will highlight some useful updates and the main differences between the two versions.
NEBUILDER^® Assembly Tool 2.0 Fragments Amplified by PCR

This video demonstrates how the NEBuilder Assembly Tool can be used to generate overlap sequences for the assembly of two fragments into a vector. In this scenario, all fragments are amplified by PCR and the vector does not have convenient restriction sites.
NEBUILDER^® Assembly Tool 2.0 Restriction Enzyme Digest

This video demonstrates how to use the NEBuilder® Assembly Tool to build a construct using a restriction enzyme digested vector and two PCR-generated inserts. The tool will help to design PCR primers containing the required overlap sequences. We will also regenerate one of the restriction enzyme recognition sites.
NEBioCalculator^® - Using the ds: mass < — > moles module to plan an NEBuilder^® HiFi DNA Assembly Reaction

This tutorial describes the use of the NEBioCalculator web tool module that converts mass to, or from, moles to help plan an NEBuilder HiFi DNA Assembly reaction