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.
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.
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
From DNA construction to protein in 1 day
Cell-free workflow combining Gibson Assembly cloning or self-circularization with rolling circle amplification using the EquiPhi29 DNA Amplification Kit.
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.
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.
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.
Benefits of GeneArt Gibson Assembly Cloning technology
GeneArt Gibson assembly EX kits are ideal for assembling multiple inserts.
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.
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.
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.
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.
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.
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 Kits | GeneArt Gibson Assembly EX Master Mix and Kits | |
|---|---|---|
| Number of steps in protocol | Single tube, One-step | Single tube, Two-step |
| Cloning Efficiency | Up to 95% | Up to 95% |
| Fragment size | 500 bp to 32 kb | 100 bp to 100 kb |
| Reaction Time(s) | From 15 minutes | 80 minutes |
| Number of Fragments | Up to 6 | Up to 15 |
| Formats | Available as a Master Mix, full kit with chemically competent or electrocompetent cells | Available as a Master Mix, full kit with chemically competent or electrocompetent cells |
| Ideal for | Simple assemblies | Complex 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 fragments | Fragment size | Length of overlap regions |
|---|---|---|
| 1-2 | ≤ 8 kb | 20–40 bp |
| 8–32 kb | 25–40 bp | |
| 3-5 | ≤ 8 kb | 40 bp |
| 8–32 kb | 40–100 bp | |
| 6+ | 100 bp to 100 kb | 50–100bp |
Options for generating DNA
| PCR amplification | GeneArt Strings DNA Fragments | GeneArt 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. Order now | 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. Order now |
Ordering information
GeneArt Gibson Assembly HiFi Kits
GeneArt Gibson Assembly EX Kits
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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.
For Research Use Only. Not for use in diagnostic procedures.