In-Fusion HD Cloning Plus
In-Fusion HD Cloning Plus
Brand: Takara Bio.
In stock
SKU
In-Fusion HD Cloning Plus
Overview
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.
- Subcloning is unnecessary: Clone any insert, into any locus, in any vector, with just one reaction
- Highly efficient: Over 95% efficiency demonstrated with a broad range of fragment sizes, from 0.5 kb to 15 kb
- Seamless construction: Final constructs have no extra base pairs left over (as is often the case with restriction digest or TA cloning)
- Flexibility to clone single or multiple fragments: Clone single or multiple DNA fragments simultaneously, in a single reaction
- Can be used to perform site-directed mutagenesis: Click here to learn how
Applications
- PCR cloning
- High-throughput cloning (see full range of reagent options on our high-throughput cloning page)
- Multiple-fragment cloning
- Gene synthesis
- Gene design
- Mutagenesis
- Domain swapping
- Domain modification
In-Fusion HD Cloning Plus system components
All In-Fusion HD Cloning products contain the In-Fusion HD Enzyme Premix and reagents for control experiments. In-Fusion HD Cloning Plus also includes:
-
In-Fusion HD Enzyme Premix
In-Fusion HD enzyme mix fuses DNA fragments efficiently and precisely by recognizing a 15-bp overlap at their ends—without the use of ligase. Provided in a liquid format as a 5X enzyme premix in 20-μl or 100-μl aliquots, depending on kit size. Note: the EcoDry kit format includes lyophilized cloning enzyme, prealiquoted in 8-well tube strips or 96-well plates, depending on kit size. -
CloneAmp HiFi PCR Premix
CloneAmp HiFi PCR Premix is a convenient 2X liquid master mix that offers exceptionally accurate, efficient, and fast DNA amplification. The premix contains dNTPs and an optimized buffer, allows rapid set-up of PCR reactions, and facilitates successful cloning. The polymerase master mix is provided in 625-μl aliquots in both EcoDry and liquid kits. -
Stellar Competent Cells
High-efficiency competent cells are essential to the success of In-Fusion Cloning. An E. coli HST08 strain is included that provides high transformation efficiency (greater than 5 x 108 cfu/µg) and complements the efficiency of all In-Fusion HD Cloning Plus kits. Cells are provided in 100-μl aliquots in individual tubes. The 96-rxn kit includes the cells prealiquoted in 96-well plates (20 μl in each well). -
Cloning Enhancer or NucleoSpin Gel and PCR Clean-up kit
Cloning Enhancer (CE) is an easy-to-use enzyme premix that removes background plasmid DNA and PCR residue, eliminating the need for PCR purification prior to cloning. In-Fusion HD Cloning Plus CE kits are ideal for cloning when there is a single PCR product on the gel, and can be used as part of a streamlined workflow for high-throughput systems. Cloning Enhancer is provided in 50-μl aliquots. Alternatively, NucleoSpin Gel and PCR Clean-up is suitable for gel extraction as well as PCR purification . Provided as individual columns or 96-well plate purifications, depending on kit size.
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).
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.
The In-Fusion cloning protocol
The In-Fusion cloning protocol.
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In-Fusion HD Cloning Plus system components
All In-Fusion HD Cloning products contain the In-Fusion HD Enzyme Premix and reagents for control experiments. In-Fusion HD Cloning Plus also includes:
-
In-Fusion HD Enzyme Premix
In-Fusion HD enzyme mix fuses DNA fragments efficiently and precisely by recognizing a 15-bp overlap at their ends—without the use of ligase. Provided in a liquid format as a 5X enzyme premix in 20-μl or 100-μl aliquots, depending on kit size. Note: the EcoDry kit format includes lyophilized cloning enzyme, prealiquoted in 8-well tube strips or 96-well plates, depending on kit size. -
CloneAmp HiFi PCR Premix
CloneAmp HiFi PCR Premix is a convenient 2X liquid master mix that offers exceptionally accurate, efficient, and fast DNA amplification. The premix contains dNTPs and an optimized buffer, allows rapid set-up of PCR reactions, and facilitates successful cloning. The polymerase master mix is provided in 625-μl aliquots in both EcoDry and liquid kits. -
Stellar Competent Cells
High-efficiency competent cells are essential to the success of In-Fusion Cloning. An E. coli HST08 strain is included that provides high transformation efficiency (greater than 5 x 108 cfu/µg) and complements the efficiency of all In-Fusion HD Cloning Plus kits. Cells are provided in 100-μl aliquots in individual tubes. The 96-rxn kit includes the cells prealiquoted in 96-well plates (20 μl in each well). -
Cloning Enhancer or NucleoSpin Gel and PCR Clean-up kit
Cloning Enhancer (CE) is an easy-to-use enzyme premix that removes background plasmid DNA and PCR residue, eliminating the need for PCR purification prior to cloning. In-Fusion HD Cloning Plus CE kits are ideal for cloning when there is a single PCR product on the gel, and can be used as part of a streamlined workflow for high-throughput systems. Cloning Enhancer is provided in 50-μl aliquots. Alternatively, NucleoSpin Gel and PCR Clean-up is suitable for gel extraction as well as PCR purification . Provided as individual columns or 96-well plate purifications, depending on kit size.
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).
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.
The In-Fusion cloning protocol
The In-Fusion cloning protocol.
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