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
- 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 12 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
Want to try NEBuilder HiFi DNA Assembly?
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NEBuilder HiFi DNA assembly offers unique features over other DNA assembly methods, including:
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 DNA Assembly Cloning Kit (NEB #E5520)
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.
|NEBuilder® Protocol Calculator
Use to generate a customized protocol for your NEBuilder HiFi DNA Assembly reaction.
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 firstname.lastname@example.org. The use of these products may require you to obtain additional third party intellectual property rights for certain applications.
- NEBuilder HiFi DNA Assembly Reaction Protocol
- NEBuilder® HiFi DNA Assembly Electrocompetent Transformation Protocol
- NEBuilder® HiFi DNA Assembly Chemical Transformation Protocol (E2621, E5520, E2623)
- NEBuilder Assembly of a PCR Fragment
- Protocol for cloning DNA containing repeat elements (C3040)
- Protocol for Bridging double-stranded DNA with a single-stranded DNA oligo using NEBuilder HiFi DNA Assembly (NEB #E2621)
- Protocol for assembling annealed DNA oligonucleotides and a double-stranded DNA vector using NEBuilder HiFi DNA Assembly (NEB #E2621)
- Bridging dsDNA with a ssDNA Oligo and NEBuilder® HiFi DNA Assembly to create an sgRNA-Cas9 Expression Vector
- 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
- Construction of an sgRNA-Cas9 expression vector via single-stranded DNA oligo bridging of double-stranded DNA fragments
- NEBuilder® Protocol Calculator
- Synthetic Biology/DNA Assembly Selection Chart
- Optimization Tips for NEBuilder® HiFi DNA Assembly and NEB® Gibson Assembly
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email email@example.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.
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.
In this webinar, we will explore some of the latest applications for NEBuilder HiFi DNA Assembly. View full Q & A summary.
Find out how NEBuilder® HiFi DNA Assembly can reliably join DNA fragments in a single tube, isothermal reaction, with advantages over NEB 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.
Learn how NEBuilder® HiFi DNA Assembly bridges dsDNA with a ssDNA oligo.
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.
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.
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
- 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