GeneArt Gibson Assembly® Cloning

Effortlessly arrange DNA fragments to create something truly epic with GeneArt Gibson Assembly Cloning

Gibson Assembly Cloning is an elegant and robust seamless or scar less cloning methodology that has been widely adopted by the scientific community and enables the assembly of multiple DNA fragments regardless of length or end compatibility in a highly efficient, seamless method.

When combined with GeneArt DNA Strings fragments or GeneArt Gene Synthesis GeneArt Gibson Assembly is the optimal choice for building large and demanding constructs.

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Product highlights

  • Elegant—Homologous overlap design allowing up to 15 DNA fragments to build the perfect seamless clone
  • Efficient—Cloning efficiencies up to >95% help save time and resources to accelerate your research
  • Proven—The Gibson Assembly method has been referenced in thousands of peer reviewed publications
  • Versatile—Can be used to build entire genomes de novo
  • Flexible—GeneArt Gibson Assembly kits are available complete with competent cells or master mix formats for maximum flexibility

Applications

  • Cloning and shotgun cloning
  • Protein Expression
  • Site-directed mutagenesis
  • Small Genome Assembly1

Featured resources

Gibson Assembly HiFi Cloning Kit video

Utilizing both homology and oligonucleotide stitching techniques to build large constructs

Listen to a scientist discuss homology and oligonucleotide stitching techniques to build large constructs.

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Gibson Assembly EX Cloning kit video

Evaluation of GeneArt Gibson Assembly EX Cloning technology to build large and complex assemblies

Listen to a scientist evaluate GeneArt Gibson Assembly EX Cloning technology to build complex assemblies.

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Gibson assembly for scarless cloning

DNA Cloning Tips—Build Clones with DNA Fragments using GeneArt Gibson Assembly Cloning kits

Building large DNA constructs that contain no extraneous sequences is often a challenging task. One of the key engineering tools designed to help in constructing these large constructs is Gibson Assembly cloning.

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Benefits of GeneArt Gibson Assembly Cloning technology

GeneArt Gibson assembly EX kits are ideal for assembling multiple inserts. 

GeneArt Gibson Assembly EX Cloning kits provide high transformation efficiency options when using larger numbers of inserts

Figure 1. GeneArt Gibson Assembly EX Cloning kits provide high transformation efficiency options when using larger numbers of inserts. Assembly of 6, 8 and 10 fragments of 0.5kb in pcDNA 3.4 transformed in Invitrogen TOP10 Competent Cells.

GeneArt Gibson Assembly HiFi kits offers a very cost effective and efficient way of assembling smaller numbers of fragments. For larger assemblies the GeneArt Gibson EX Master Mixes and Kits are available.

GeneArt Gibson Assembly HiFi kits provide high cloning efficiency using a single insert to multiple insert designs

Figure 2. GeneArt Gibson Assembly HiFi kits provide high cloning efficiency using a single insert to multiple insert designs. Assembly of 1, 2 and 4 - 1kb fragments in pCDNA 3.4 using TOP10 competent cells. GeneArt Gibson Assembly HiFi kits are the most cost-effective method and time-saving method for building large assemblies, particularly when used with GeneArt Strings DNA Fragments or 100% sequenced, GeneArt Gene Synthesis.

Both GeneArt Gibson Assembly HiFi and EX technologies are available in master mix formats, complete kits and with a choice of chemically competent and electrocompetent cells which both offer high transformation efficiencies.

GeneArt Gibson Assembly EX Cloning kits provide high transformation efficiency options when using larger numbers of inserts

Figure 3. GeneArt Gibson Assembly EX Cloning kits provide high transformation efficiency options for high cloning efficiency. Assembly of 5 inserts of 2kb in pUC19L using Invitrogen TOP10 chemically competent cells and ElectroMAX DH10B electrocompetent cells.

GeneArt Gibson assembly EX kits are ideal for assembling multiple inserts. 

GeneArt Gibson Assembly EX Cloning kits provide high transformation efficiency options when using larger numbers of inserts

Figure 1. GeneArt Gibson Assembly EX Cloning kits provide high transformation efficiency options when using larger numbers of inserts. Assembly of 6, 8 and 10 fragments of 0.5kb in pcDNA 3.4 transformed in Invitrogen TOP10 Competent Cells.

GeneArt Gibson Assembly HiFi kits offers a very cost effective and efficient way of assembling smaller numbers of fragments. For larger assemblies the GeneArt Gibson EX Master Mixes and Kits are available.

GeneArt Gibson Assembly HiFi kits provide high cloning efficiency using a single insert to multiple insert designs

Figure 2. GeneArt Gibson Assembly HiFi kits provide high cloning efficiency using a single insert to multiple insert designs. Assembly of 1, 2 and 4 - 1kb fragments in pCDNA 3.4 using TOP10 competent cells. GeneArt Gibson Assembly HiFi kits are the most cost-effective method and time-saving method for building large assemblies, particularly when used with GeneArt Strings DNA Fragments or 100% sequenced, GeneArt Gene Synthesis.

Both GeneArt Gibson Assembly HiFi and EX technologies are available in master mix formats, complete kits and with a choice of chemically competent and electrocompetent cells which both offer high transformation efficiencies.

GeneArt Gibson Assembly EX Cloning kits provide high transformation efficiency options when using larger numbers of inserts

Figure 3. GeneArt Gibson Assembly EX Cloning kits provide high transformation efficiency options for high cloning efficiency. Assembly of 5 inserts of 2kb in pUC19L using Invitrogen TOP10 chemically competent cells and ElectroMAX DH10B electrocompetent cells.

Which GeneArt Gibson Assembly Technology should I choose?

 GeneArt Gibson Assembly HiFi Master Mix and KitsGeneArt Gibson Assembly EX Master Mix and Kits
Number of steps in protocolSingle tube, One-stepSingle tube, Two-step  
Cloning EfficiencyUp to 95%Up to 95%
Fragment size500 bp to 32 kb100 bp to 100 kb
Reaction Time(s)From 15 minutes80 minutes
Number of FragmentsUp to 6Up to 15
FormatsAvailable as a Master Mix, full kit with chemically competent or electrocompetent cellsAvailable as a Master Mix, full kit with chemically competent or electrocompetent cells
Ideal forSimple assembliesComplex assemblies, and projects using large fragments  

GeneArt Gibson Assembly technology relies on homologous recombination to assemble adjacent DNA fragments sharing end-terminal homology. The optimal length of the homologous fragment ends region depends on the number and length of the fragments in the assembly reaction. The following table lists the suggested length of the end-terminal homology.

Number of fragmentsFragment sizeLength of overlap regions
1-2≤ 8 kb20–40 bp
8–32 kb25–40 bp
3-5≤ 8 kb40 bp
8–32 kb40–100 bp
6+100 bp to 100 kb50–100bp

Options for generating DNA

PCR amplificationGeneArt Strings and High Q Strings DNA FragmentsGeneArt Gene Synthesis
Generate PCR fragments with vector homology or take archived PCR fragments and use primer stitching for Gibson Assembly. And with a polymerase like Platinum SuperFi II you get unparalleled fidelity.

Get more information on PCR Products
GeneArt Strings DNA Fragments are custom-made, uncloned, double-stranded linear DNA fragments assembled from synthetic oligonucleotides using the same high-quality process developed for Invitrogen GeneArt Gene Synthesis. For even greater sequence accuracy GeneArt High-Q Strings offer a leading accuracy in the market with an error rate of less than 1 in 10,000 bp.

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GeneArt Gene Synthesis clones are 100% sequence verified. Simply excise from our standard cloning vector (pMX) and you’re ready to start building error-free assemblies. And with our superSPEED gene synthesis service you can get error free fragments even faster.

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Ordering information

GeneArt Gibson Assembly HiFi Kits

GeneArt Gibson Assembly EX Kits

1Enzymatic assembly of DNA molecules up to several hundred kilobases. Gibson et. al., Nat Methods. 2009 May; 6(5):343-5

Gibson Assembly® is a registered trademark of SGI-DNA, Inc. used under permission and license.
NEBuilder® is a registered trademarks of New England Biolabs, Inc.
In-Fusion® is a registered trademarks of Takara Bio USA, Inc.