NEBuilder® HiFi DNA Assembly – Benefits Over NEB Gibson Assembly®

Return to NEBuilder® HiFi DNA Assembly

NEBuilder HiFi DNA Assembly offers several advantages over NEB Gibson Assembly. 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 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.)
  • 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 Gibson Assembly-designed (and other) fragments
  • NEBuilder Assembly Tool is available to help with primer design
  • 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)


Performance Data

See how NEBuilder HiFi outperforms NEB Gibson Assembly.

Improved efficiency and accuracy

Improved fidelity


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. .

GIBSON ASSEMBLY® is a registered trademark of Synthetic Genomics, Inc.