NEBuilder® HiFi DNA Assembly – Benefits Over In-Fusion® HD

Return to NEBuilder® HiFi DNA Assembly

NEBuilder HiFi DNA Assembly offers several advantages over In-Fusion HD. These include: higher accuracy due to the use of a high-fidelity polymerase, the ability to assemble both 5´- and 3´-end mismatches, lower DNA input requirements and the ability to bridge two dsDNA fragments with a ssDNA oligo (data not shown). NEBuilder HiFi DNA Assembly is the clear choice for efficient and accurate DNA assembly.


  • Enjoy less screening/re-sequencing of constructs, with virtually error-free, high-fidelity assembly
  • Join DNA fragments together more efficiently, even with larger fragments or low DNA inputs
  • Use NEBuilder HiFi in successive rounds of assembly, because it removes 5´- and 3´-end mismatches (Save time by avoiding time-consuming PCR amplification steps.)
  • Product is covalently sealed and will not fall apart during PCR
  • Bridge two ds-fragments with a synthetic ssDNA oligo for simple and fast construction (e.g., linker insertion or gRNA library)
  • Switch from other systems easily, as NEBuilder HiFi is compatible with In-Fusion-designed (and other) fragments
  • NEBuilder Assembly Tool is available to help with primer design
  • No licensing fee requirements from NEB for NEBuilder products
  • Choose from: NEBuilder HiFi DNA Assembly Cloning Kit (NEB #E5520), NEBuilder HiFi DNA Assembly Bundle for Large Fragments (NEB #E2623)and NEBuilder HiFi DNA Assembly Master Mix (NEB #E2621)

"I want to tell you that NEBuilder works really well! I tried to do the installation first with Takara's InFusion cloning, and I got no positive colonies...After a hard look at my procedure, I couldn't figure out what I did wrong. Not finding a technical mistake, I repeated the exact same experiment (in terms of molar ratios and nanograms of DNA used) except with the NEBuilder free sample (thank you for that!). I got positive colonies and my insert is now sent off for sequencing! Five stars! *****”

–  J.P., Ph.D. Candidate
University of Iowa, College of Pharmacy, Division of Medicinal and Natural Products Chemistry

Performance Data

See how NEBuilder HiFi outperforms In-Fusion.

Two-fragment assembly with and without 5´- and 3´-end mismatches

Four-fragment assembly

Oligo & vector 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 The use of these products may require you to obtain additional third party intellectual property rights for certain applications.

IN-FUSION® is a registered trademark of ClonTech Laboratories, Inc.