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  • ‹ Back to In-Fusion Cloning tech notes
  • In-Fusion Cloning of sgRNAs
  • Sequence accuracy in seamless cloning
  • Rapid, high-throughput cloning for antibody development
  • De novo insertion of small fusion protein tags
  • Improving background over Gibson Assembly
  • A successful alternative to ligation cloning
  • Single- and multiple-insert cloning
  • Direct cloning into large vectors
  • Easy cloning into lentiviral vectors
  • Outperforming TOPO cloning
  • Solve a synthesis challenge with easy multiple-insert cloning
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Home › Learning centers › Cloning › In-Fusion Cloning tech notes › Sequence accuracy in seamless cloning

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  • In-Fusion Cloning tech notes
    • In-Fusion Cloning of sgRNAs
    • Sequence accuracy in seamless cloning
    • Rapid, high-throughput cloning for antibody development
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    • Improving background over Gibson Assembly
    • A successful alternative to ligation cloning
    • Single- and multiple-insert cloning
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    • Solve a synthesis challenge with easy multiple-insert cloning
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Tech Note

Higher sequence accuracy for seamless constructs with In-Fusion Cloning

  • Seamless cloning enables directional, multiple-insert cloning without scar sequences
  • Methods for seamless cloning vary in their enzymatic mechanisms
  • Sequence accuracy is improved with ligase-free In-Fusion Cloning compared to Gibson Assembly and NEBuilder kits
Introduction Results Conclusions Methods References

Introduction  

Researchers have several options when it comes to seamless cloning methods. The mechanism for each varies quite a bit, as does the resulting sequence accuracy, which we will discuss below. The benefits to any seamless method include the lack of unwanted nucleotides—or scar sequences—between vector and insert, as well as the ability to clone multiple inserts in a specific order in just one round of cloning. However, these research tools lose some of their power if accuracy and/or efficiency suffer. High-throughput applications are particularly affected by any pain points in this regard, as problems with potential error rates or background levels will be magnified when projects are scaled up to accommodate larger cloning needs.

Several commercially available seamless cloning kits, including Gibson Assembly and NEBuilder, utilize a three-enzyme mix consisting of an exonuclease, polymerase, and ligase (Gibson et al. 2009). The mechanism for In-Fusion Cloning (Irwin et al. 2012) differs from this process in a simple but significant way. Both mechanisms require that inserts are amplified by PCR such that they include homologous overlaps with a linear vector or other adjacent fragments, and both utilize an exonuclease to chew back the DNA of linear fragments to create cohesive sticky ends. At this point, the workflows part ways. With the three-enzyme methods, a polymerase fills in any nucleotide gaps, and a DNA ligase then seals the nicks in the annealed fragments prior to the transformation step. This process runs the risk of introducing sequence errors and mismatches at the cloning junction. In contrast, In-Fusion technology does not use a polymerase nor DNA ligase in the cloning mechanism, thus eliminating the potential for nucleotide errors. Cohesive bonds are formed in vivo, preserving the sequence integrity of the clone. In an effort to understand the repercussions of the differences in these mechanisms, we used commercially available kits for each to perform the demanding task of multiple-insert cloning, specifically looking at sequence accuracy as a measure of success.

Results  

In the experiment below, we compared two kits for the three-enzyme method (NEBuilder DNA HiFi Assembly from NEB and the Gibson Assembly Kit from SGI) with the liquid and lyophilized (EcoDry) versions of Takara Bio's In-Fusion Cloning HD Plus kits in order to assess the sequence accuracy of clones made with each method. Each seamless cloning kit was used to clone five inserts (947 bp, 717 bp, 697 bp, 405 bp, and 1,005 bp) into a 2.7-kb vector backbone, with the molar ratio of vector to insert set at 1:2 (Table I).

Table I. Cloning reaction setup to achieve a vector-to-insert molar ratio of 1:2
Linear fragment Amount
Insert 1 (947 bp) 40 ng
Insert 2 (717 bp) 40 ng
Insert 3 (697 bp) 32 ng
Insert 4 (405 bp) 20 ng
Insert 5 (1,005 bp) 45 ng
Vector (2,671 bp) 37 ng

All clones were designed with 20-bp homologous overlaps between each pair of linear fragments, as per the recommendations for multiple-insert reactions with In-Fusion Cloning. Reactions for each system were carried out per their respective manufacturer's instructions, and 1/10 dilutions were plated on selective medium. 20 clones from each kit were chosen at random in order to check sequence fidelity. Both three-enzyme methods yielded clones with an average of 80% accuracy, while In-Fusion Cloning produced a higher level of accuracy (Table II), with liquid and EcoDry kits yielding 90% and 100%, respectively.

Table II. Sequence accuracy comparison for all four seamless cloning kits
Commercial kit Sequence accuracy (average of 20 clones)
In-Fusion HD Cloning Plus (liquid format) 90%
In-Fusion HD EcoDry Cloning Plus (lyophilized format) 100%
NEBuilder HiFi DNA Assembly Cloning Kit (NEB) 80%
Gibson Assembly Kit (SGI) 80%

Conclusions  

The sequence accuracy of In-Fusion Cloning is consistently higher than both commercial kits for three-enzyme cloning methods, regardless of which format of the In-Fusion enzyme mix is used. It is likely that this difference stems from the lack of ligase and polymerase in the In-Fusion Cloning mix, as both ligase and polymerase performance can be error-prone. Sequence errors can be especially problematic in multiple-insert cloning, as there are more cloning junctions where nucleotide mismatches can occur. The reliable results provided by In-Fusion technology make it especially adaptable to high-throughput workflows. Higher accuracy means less screening and fewer missed clones in large batches, which would then require additional rounds of cloning to recover.

Methods  

Each seamless cloning kit was used according to the manufacturer's instructions. Vector-to-insert molar ratios were set at 1:2 for each reaction. Reactions for each kit were run in triplicate, with the exception of the liquid In-Fusion Cloning kit, which was performed once to reproduce previous data. NEB 5-alpha Competent E. coli (High Efficiency; NEB) were transformed with the cloning reactions from the NEBuilder HiFi DNA Assembly Cloning Kit (NEB), TransforMax EPI300 Electrocompetent and Chemically Competent E. coli were transformed with the cloning reactions from the Gibson Assembly Kit (SGI), and Stellar Competent Cells (included in the In-Fusion Cloning kits) were transformed with the cloning reactions from both In-Fusion HD Cloning Plus and In-Fusion HD EcoDry Cloning Plus. 1/10 dilutions were plated on selective medium for each transformation. 20 clones were chosen at random from transformation plates of each cloning kit and checked for sequence accuracy via Sanger sequencing.

References  

Gibson D.G., et al. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nature Methods 6(5): 343–345 (2009).

Irwin C.R., Farmer A., Willer D.O., Evans D.H. In-Fusion Cloning with Vaccinia Virus DNA Polymerase. Vaccinia Virus and Poxvirology (Methods and Protocols). 890:23–35 (2012).

Related Products

Cat. # Product Size Price License Quantity Details
638920 In-Fusion® HD Cloning Plus 96 Rxns $1457.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD Cloning Plus kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. In addition to the cloning kit, this package includes: a NucleoSpin 96 PCR Clean-Up Kit, which can be used for spin-column purification of PCR products; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638911 In-Fusion® HD Cloning Plus 100 Rxns $1514.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD Cloning Plus kit enables high-efficiency, high-fidelity directional cloning of one or more PCR fragments into any vector. In addition to the cloning kit, this package includes: a NucleoSpin Gel and PCR Clean-Up Kit, which can be used for spin-column purification of PCR products; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638910 In-Fusion® HD Cloning Plus 50 Rxns $892.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD Cloning Plus kit enables high-efficiency, high-fidelity directional cloning of one or more PCR fragments into any vector. In addition to the cloning kit, this package includes: a NucleoSpin Gel and PCR Clean-Up Kit, which can be used for spin-column purification of PCR products; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638909 In-Fusion® HD Cloning Plus 10 Rxns $231.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD Cloning Plus kit enables high-efficiency, high-fidelity directional cloning of one or more PCR fragments into any vector. In addition to the cloning kit, this package includes: a NucleoSpin Gel and PCR Clean-Up Kit, which can be used for spin-column purification of PCR products; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

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The In-Fusion cloning protocol.
638919 In-Fusion® HD Cloning Plus CE 96 Rxns $1496.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD Cloning Plus CE kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. In addition to the cloning kit, this package includes: Cloning Enhancer, which removes background plasmid DNA and PCR residue, eliminating the need for PCR insert purification prior to cloning; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

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The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

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The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638918 In-Fusion® HD Cloning Plus CE 100 Rxns $1567.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD Cloning Plus CE kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. In addition to the In-Fusion HD Cloning Kit, this package includes: Cloning Enhancer, which removes background plasmid DNA and PCR residue, eliminating the need for PCR insert purification prior to cloning; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638917 In-Fusion® HD Cloning Plus CE 50 Rxns $948.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD Cloning Plus CE kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. In addition to the In-Fusion HD Cloning Kit, this package includes: Cloning Enhancer, which removes background plasmid DNA and PCR residue, eliminating the need for PCR insert purification prior to cloning; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

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The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638916 In-Fusion® HD Cloning Plus CE 10 Rxns $238.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD Cloning Plus CE kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. In addition to the In-Fusion HD Cloning Kit, this package includes: Cloning Enhancer, which removes background plasmid DNA and PCR residue, eliminating the need for PCR insert purification prior to cloning; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638915 In-Fusion® HD EcoDry™ Cloning Plus 96 Rxns $1449.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD EcoDry Cloning Plus kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. The EcoDry Kit provides In-Fusion enzyme in a convenient, lyophilized (dry) format. In addition to the cloning kit, this package includes: a NucleoSpin Gel and PCR Clean-Up Kit, which can be used for both gel extraction and spin-column purification of PCR products; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638914 In-Fusion® HD EcoDry™ Cloning Plus 48 Rxns $1020.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD EcoDry Cloning Plus kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. The EcoDry Kit provides In-Fusion enzyme in a convenient, lyophilized (dry) format. In addition to the cloning kit, this package includes: a NucleoSpin Gel and PCR Clean-Up Kit, which can be used for both gel extraction and spin-column purification of PCR products; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638913 In-Fusion® HD EcoDry™ Cloning Plus 24 Rxns $518.00

License Statement

ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD EcoDry Cloning Plus kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. The EcoDry Kit provides In-Fusion enzyme in a convenient, lyophilized (dry) format. In addition to the cloning kit, this package includes: a NucleoSpin Gel and PCR Clean-Up Kit, which can be used for both gel extraction and spin-column purification of PCR products; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

Notice to purchaser

Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

Documents Components You May Also Like Image Data Resources

Back

The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

Back

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

Back

The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.
638912 In-Fusion® HD EcoDry™ Cloning Plus 8 Rxns $210.00

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ID Number  
28 This product is covered by the claims of U.S. Patent No. 7,575,860 and its foreign counterpart patent claims.

The In-Fusion HD EcoDry Cloning Plus kit enables high-efficiency, high-fidelity, directional cloning of one or more PCR fragments into any vector. The EcoDry Kit provides In-Fusion enzyme in a convenient, lyophilized (dry) format. In addition to the cloning kit, this package includes: a NucleoSpin Gel and PCR Clean-Up Kit, which can be used for both gel extraction and spin-column purification of PCR products; Stellar Competent Cells, an E. coli HST08 strain that provides high transformation efficiency paired with blue-white screening capability when used with pUC plasmid vectors; and CloneAmp HiFi PCR Premix, a convenient high-fidelity PCR 2X master mix that provides exceptionally accurate and efficient DNA amplification.

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Our products are to be used for Research Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval.

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The In-Fusion HD Cloning System has been optimized for cloning large fragments

The In-Fusion HD Cloning System has been optimized for cloning large fragments
The In-Fusion HD Cloning System has been optimized for cloning large fragments. Ten out of ten colonies contain the correct insert (100% efficiency) when cloning fragments as large as 15 kb (results confirmed by colony PCR screening).

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The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction

The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction
The In-Fusion HD Cloning System has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.

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The In-Fusion cloning protocol

The In-Fusion cloning protocol
The In-Fusion cloning protocol.


In-Fusion Cloning for high-throughput antibody development

A rapid, high-throughput cloning workflow for antibody development

Generating monoclonal antibodies requires efficient and rapid protocols for cloning, expression, and screening. Traditional cloning creates bottlenecks in this research, as it is often hampered by lengthy protocols, low efficiency, and reading frame errors. See how one lab has streamlined the process with In-Fusion technology, a highly accurate, seamless cloning method which is easily adaptable to high-throughput workflows.

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Takara Bio USA, Inc. (TBUSA, formerly known as Clontech Laboratories, Inc.) provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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Capturem Trypsin for a rapid, efficient mass spectometry workflow at room temperature.

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Capturem Trypsin provides rapid, efficient, and complete digestion of protein samples, allowing an uninterrupted mass spectometry workflow at room temperature for downstream protein analysis. This product utilizes our novel Capturem technology in a spin column format with membrane-immobilized trypsin. Capturem Trypsin Columns may be used to completely digest protein samples in less than a minute with digestion efficiencies (protein coverage) comparable to or better than those obtained using in-solution trypsin digestion.

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Takara Bio USA, Inc. (TBUSA, formerly known as Clontech Laboratories, Inc.) provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, TBUSA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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