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 [email protected]. 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.