Product Class: Kit

Gibson Assembly® Master Mix

Gibson Assembly Master Mix (NEB #M5510) has been reformulated with components containing Recombinant Albumin (rAlbumin) beginning with Lot #10229799. Learn more.

Have you tried NEBuilder HiFi DNA Assembly? NEBuilder HiFi offers several advantages over NEB Gibson Assembly. For more information, visit NEBuilderHiFi.com.

Product Introduction

Gibson Assembly allows for successful assembly of multiple DNA fragments, regardless of fragment length or end compatibility.

  • Assembly and transformation in just under two hours
  • Flexible sequence design (scar-less cloning)
  • No PCR clean-up step required
  • High transformation efficiencies for inserts up to 20 kb
  • Easily adapted for multiple DNA manipulations, including site-directed mutagenesis
Catalog # Size Concentration
E2611S 10.0 reactions
E2611L 50.0 reactions

Featured Videos

View Video Library
  • GibsonOverviewVideo_thumb

    Introduction to Gibson Assembly®

  • FA_Fig4_GibsonVideo_thumb

    Gibson Assembly Workflow

  • NEBTVToolsThumbnailNEBuilderAssemblyTool20WhatsNew1920

    NEBUILDER® Assembly Tool 2.0 What’s New?

  • NEBTVToolsThumbnailNEBuilderAssemblyTool20Fragments Amplified by PCR

    NEBUILDER® Assembly Tool 2.0 Fragments Amplified by PCR

  • NEBTVToolsNEBuilderAssemblyTool20RestrictionEnzymeDigest1920

    NEBUILDER® Assembly Tool 2.0 Restriction Enzyme Digest

  • PrimerDesignFragmentAssembly_720

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

Protocols, Manuals & Usage

Protocols

  1. Gibson Assembly® Master Mix – Assembly (E2611)
  2. Gibson Assembly® Chemical Transformation Protocol (E2611)
  3. Gibson Assembly® Electrocompetent Cells Transformation Protocol (E2611)

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

Application Notes

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. I am not sure whether to choose NEBuilder HiFi DNA Assembly or NEB Gibson Assembly? How are the products different?
  2. What are the advantages of this method compared to traditional cloning methods?
  3. How large a DNA fragment can I assemble?
  4. How many fragments of DNA can be assembled in one reaction?
  5. What are the shortest overlaps that can be used with this assembly method?
  6. What are the longest overlaps that can be used with this method?
  7. Can ≤ 200 bp dsDNA fragments be assembled by this method?
  8. Can ssDNA oligonucleotides be combined and assembled with dsDNA fragments?
  9. Can longer or shorter incubation times be used?
  10. Will the reaction work at other temperatures?
  11. Is it necessary to inactivate restriction enzymes after vector digestion?
  12. I would like to produce overlapping dsDNA fragments by PCR. Do I need to use PCR primers that have been purified by PAGE or HPLC?
  13. I would like to assemble ssDNA oligonucleotides into dsDNA fragments. Do I need to use oligonucleotides that have been purified by PAGE or HPLC?
  14. Can I use a 15-nt overlap that is entirely composed of His-tag repeats (i.e. CACCACCACCACCAC)?
  15. Can you PCR-amplify the assembled product?
  16. What should I do if my assembly reaction yields no colonies, a small number of colonies, or clones with the incorrect insert size following transformation into E. coli?
  17. How can I reduce the number of vector-only background colonies?
  18. What type of competent cells are suitable for transformation of DNA constructs created using Gibson Assembly?
  19. Can I use electroporation instead of chemical transformation?
  20. Are there any differences between the Gibson Assembly Master Mix (NEB #E2611) and Gibson Assembly Master Mix included in the Gibson Assembly Cloning Kit (NEB #E5510)?
  21. Are there any differences between the requirements for 2-3 fragmentassemblies versus 4–6?
  22. The Gibson Assembly Master Mix control reaction is not giving me any colonies. Why?
  23. When using a polymerase that doesn't contain a 3'-5' exonuclease activity (such as Taq DNA Polymerase) to amplify fragments to be used in a Gibson Assembly reaction, should I be concerned about the potential 3' mismatch generated by the addition of a non-templated nucleotide?
  24. Is storing Gibson Assembly Master Mix at -80°C harmful?
  25. I would like to use NEBuilder but am concerned about user data privacy. How does NEB handle the information that I enter into NEBuilder?
  26. Can I Use other primer design tools such as SnapGene for Gibson Assembly, to design primers for NEBuilder HiFi DNA Assembly?
  27. What antibiotic resistance is encoded in the NEBuilder Positive Control (assembly control)?

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