Push the Limits of Golden Gate Assembly
20+ fragment DNA assembly achievable with high efficiency and accuracy!

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With constant advances both in the development of new enzymes (e.g., BsaI-HFv2) and research on maximizing enzyme functionality (e.g., ligase fidelity), NEB has become the industry leader in pushing the limits of Golden Gate Assembly (and related methods such as MoClo, GoldenBraid, Mobius Assembly and Loop Assembly). NEB has all the products and information you need to perform complex assemblies, as demonstrated with 20+ fragment assemblies exhibiting high efficiencies and >90% accuracy, along with low backgrounds.

 

GG_LP_techNote_icon Golden Gate Assembly Workflow Diagram for complex assemblies GG_LP_GoldenGateTool_icon 

 

Golden Gate Assembly Workflow for complex assemblies

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Advances in ligase fidelity

Research at NEB has led to an increased understanding of ligase fidelity, including the development of a comprehensive method for profiling end-joining ligation fidelity in order to predict which overhangs have improved fidelity (1). Their research has enabled complex fragment assemblies with high efficiencies and >90% accuracy.  Use Ligase Fidelity Viewer to visualize overhang ligation preferences for T4 DNA ligase under various experimental conditions.

 

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Type IIS restriction enzymes for Golden Gate Assembly (BsaI-HFv2 and Esp3I)

NEB offers more Type IIS (i.e., recognize asymmetric DNA sequences and cleave outside of their recognition sequence) restriction enzymes than any other supplier, many of which are used in Golden Gate Assembly. NEB is pleased to introduce two new restriction enzymes for use in Golden Gate: Esp3I, an isoschizomer of BsmBI that is recommended for use at 37°C and is supplied with CutSmart® Buffer, and the improved BsaI-HFv2, which is optimized for Golden Gate Assembly.

For information about other Type IIS restriction enzymes used in Golden Gate Assembly, such as BbsI, BbsI-HF and SapI, view our Type IIS restriction enzyme table.

For a limited time, try a sample of BsaI-HFv2 and see how it works for you!

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1. Potapov, V. et. al. “Comprehensive Profiling of Four Base Overhang Ligation Fidelity by T4 DNA Ligase and Application to DNA Assembly.” ACS Synth. Biol. 2018. DOI: 10.1021/acssynbio.8b00333.

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.

Featured products:

NEBNext_portal_product_icon NEB® Golden Gate Assembly Kit (BsaI-HF®v2)
Contains an optimized mix of BsaI-HFv2 (optimized for Golden Gate Assembly) and T4 DNA Ligase. Can direct the accurate assembly of 2 – 20+ inserts/modules using the Golden Gate approach.
NEBNext_portal_product_icon Esp3I
An isoschizomer of BsmBI that is recommended for use at 37°C and is supplied with CutSmart Buffer, adding flexibility to your reaction setup
NEBNext_portal_product_icon BsaI-HFv2
New version of BsaI-HF that is optimized for Golden Gate Assembly.

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Videos

  1. GoldenGateAssemblyWorkflow_720

    Golden Gate Assembly Workflow

    Find out how Golden Gate Assembly can be used to quickly join multiple DNA fragments.

  2. KuceraBecky

    Restriction Enzymes in Golden Gate Assembly

    Type IIS restriction enzymes have both recognition and binding sites, but cut downstream of the recognition site, creating 4-base overhangs ideal for re-assembly. View a list of TypeIIS enzymes.