Gibson Assembly & NEBuilder HiFi DNA Assembly Header

NEBuilder HiFi DNA Assembly

NEBuilder HiFi DNA Assembly enables virtually error-free joining of DNA fragments, even those with 5´ and 3´ restriction enzyme mismatches.

This flexible kit enables simple and fast Seamless Cloning utilizing a new proprietary high-fidelity polymerase.

Find out why NEBuilder HiFi is the next generation of DNA assembly and cloning.

Order a free NEBuilder HiFi DNA Assembly Sample

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NEBuilder HiFi DNA Assembly Workflow

Illustration NEBuilder Method

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Illustration NEBuilder Workflow

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NEBuilder HiFi DNA Assembly Methode
NEBuilder & Gibson Assembly Methode Overview

Benefits over Gibson Assembly® Master Mix

  • 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
  • Bridge two double-stranded fragments with a synthetic single-stranded DNA oligo for simple and fast construction (e.g. linker insertion or gRNA libraries)
  • Switch from other systems easily, because NEBuilder HiFi is compatible with Gibson Assembly-designed (and other) fragments
  • Save time with simple and fast Seamless Cloning
  • No licensing fee requirements from NEB for the NEBuilder products

NEBuilder Assembly Tool

Need help designing primers for DNA Assembly? Try NEBuilder Assembly Tool!

NEBuilderPlease click the icon.

NEBioCalculator

Use this tool for your scientific calculations and conversions for DNA and RNA

NEBioCalculator150Please click the icon.

Comparison NEBuilder and Gibson Assembly

NEBuilder HiFi DNA Assembly Master Mix offers improved efficiency and accuracy over Gibson Assembly with different numbers of fragments.

Reactions were set up in a 2- and 6- fragment assembly reaction according to recommended reaction conditions.

NEBuilder HiFi DNA Assembly vs Gibson Assembly
NEBuilder HiFi DNA Assembly vs. Gibson Assembly with low input

NEBuilder HiFi DNA Assembly Master Mix offers improved efficiency and accuracy over Gibson Assembly, with lower amounts of DNA by increasing overlaps.

Reactions were set up in a 4- fragment assembly reaction according to recommended reaction conditions. Amount of DNA and size of overlap is shown.

NEBuilder HiFi DNA Master Mix offers improved fidelity over Gibson Assembly Master Mix

Fidelity of assembled products was compared between NEBuilder HiFi DNA Assembly Master Mix (NEB #E2621) and Gibson Assembly Master Mix (NEB #E2611). Experiments were performed using various fragment and vector sizes following suggested protocols. Experiments D and E did not yield any colonies with Gibson Assembly. Experiments H and J were performed with fragments containing 3´ restriction enzyme mismatches. Experiments B through G vary because sequences of fragments are different

NEBuilder HiFi Assembly vs. Gibson Assembly - Fidelity

Videos

Introduction to NEBuilder HiFi DNA Assembly

Primer Design and Fragment Assembly Using NEBuilder HiFi DNA Assembly or Gibson Assembly

NEBuilder HiFi DNA Assembly: Bridging dsDNA with a ssDNA Oligo

Product table

PRODUCT Prod. # SIZE PRICE
NEBuilder® HiFi DNA Assembly Cloning Kit E5520S 10 rxns 192 € In den Shop wechseln Produktinformationen
NEBuilder® HiFi DNA Assembly Master Mix E2621S 10 rxns 162 € In den Shop wechseln Produktinformationen
E2621L 50 rxns 648 € In den Shop wechseln Produktinformationen
E2621X 250 rxns 2589 € In den Shop wechseln Produktinformationen
NEBuilder® HiFi DNA Assembly Bundle for Large Fragments E2623S 20 rxns 498 € In den Shop wechseln Produktinformationen

As of: 01.01.2021

Further information can be found in our Technical Resources section or at neb.com. Information on trademarks can be found here.