NEBuilder HiFi DNA Assembly Master Mix from New England Biolabs

NEBuilder HiFi DNA Assembly Master Mix was developed to improve the efficiency and accuracy of DNA assembly. This method allows for seamless assembly of multiple DNA fragments, regardless of fragment length or end compatibility. This method has been used to assemble either single-stranded oligonucleotides or different sizes of DNA fragments with varied overlaps (15–80 bp).

The reaction includes different enzymes that work together in the same buffer -

• The exonuclease creates single-stranded 3´ overhangs that facilitate the annealing of fragments that share complementarity at one end (the overlap region)
• The polymerase fills in gaps within each annealed fragment
• The DNA ligase seals nicks in the assembled DNA

Benefits over other Assembly Master Mixes:

• 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´ restriction enzyme mismatches. (Save time by avoiding time-consuming PCR amplification steps)
• Bridge two ds-fragments with a synthetic ss-DNA oligo for simple and fast construction (e.g. linker insertion or gRNA library)
• Switch from other systems easily, because NEBuilder HiFi is compatible with other fragments.
• No licensing fee requirements from NEB for the NEBuilder products.