Comparison of DNA Assembly Reaction Types

Return to NEBuilder® HiFi DNA Assembly

NEBuilder HiFi DNA Assembly offers error-free assembly that can be used for a wide range of reaction types. Furthermore, there are no licensing fee requirements from NEB for NEBuilder HiFi DNA Assembly products. See how it compares to GeneArt® Gibson Assembly® and In-Fusion® Snap Assembly.


NEBuilder HiFi DNA Assembly GeneArt Gibson Assembly In-Fusion Snap Assembly
Assembly reaction types Assembly efficiency Covalently sealed?* Assembly efficiency Covalently sealed?* Assembly efficiency Covalently sealed?*
2-fragment assembly
No mismatch +++ Yes ++ Yes ++ No
3´- & 5´-end mismatch +++ Yes ++ Yes X No
4-fragment assembly
15-bp overlap
& no mismatch
+++ Yes ++ Yes ++ No
25-bp overlap
& no mismatch
+++ Yes ++ Yes ++ No
Oligo assembly
3´- and 5´-overhang +++ Yes ++ Yes X No
Blunt end
& no mismatch
+++ Yes ++ Yes X No
ssOligo & vector +++ Yes NP Yes X No

* Assembled products are treated with T5 exonuclease followed by PCR. Only covalently sealed products resistant to T5 exonuclease digestion can serve as templates for PCR and yield PCR product.

+++     Performs  best; recommended
++       Performs well; but other product(s) perform better
+         Performs, but  not recommended
X         Does  not perform
NP      Experiment  not performed


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