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 [email protected]. 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.