In-Fusion® HD Cloning Plus PCR Cloning Kits
PCR Cloning Kit-Seamless DNA Cloning with In-Fusion HD Cloning Plus
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Great. I would recommend to everyone who's cloning doesn't work.
Molecular biology
InFusion HD cloning system offers seamless way for rapid plasmid construction. It enables direct cloning of PCR products to linearized vectors without the need of previous purification steps or vector dephosphorylation. Definitely a useful alternative to ligation-dependent cloning.
Review Date: 25 Mar 2022 | Takara Bio
Easy to use
Mutant deletion cloning
Makes design and workload much easier. Really like the online tool.
Review Date: 12 Dec 2017 | Takara Bio
Great cloning strategy
Molecular Biology
This kit turns out to be very easy to use. It saved the cloning of a difficult and long cDNA that we needed and couldn't clone with regular methods.
Review Date: 6 Feb 2017 | Takara Bio
Great results with the In-fusion cloning kit
Plasmid cloning
It was very simple and effective to use. It’s easy to design and clone. It works like a magic mixture.
Review Date: 12 Jul 2016 | Takara Bio
Cloning
This kit is perfect for cloning larger fragment up to 4kb in first attempt. I am using this kit from last year and it’s perfect for cloning larger construct. The sensitivity is amazing and efficiency of positive clones is above 70%. I highly recommend because of its capability and sensitivity, saved lot of time as compared to normal cloning and decreases repetitive experiment as indirectly save cost as well. When I started using it was new to me as I failed with other cloning product at last I came up with this kit and worked very well since.
Review Date: 13 Oct 2015 | Takara Bio
Gateway-cloning of BCR constructs
Cloning on the principle of a fire-and-forget system. Fast and easy application in combination with efficiencies of <90% positive clones. Worth to change....
Review Date: 21 May 2015 | Takara Bio
The best for cloning large sequence inserts.
Review Date: 9 Aug 2008 | Takara Bio
PCR Cloning Kit-Seamless DNA Cloning with In-Fusion HD Cloning Plus
In-Fusion PCR Cloning Kits enable sequence independent, directional, and seamless cloning of ANY PCR fragment-or multiple fragments-into ANY linearized vector at ANY locus in a single 15-minute reaction. No additional treatment of the PCR fragment is required (such as restriction digestion, ligation, phosphorylation, or blunt-end polishing). The efficiency of In-Fusion HD Cloning Kits is over 95% across a broad insert size range (0.5 to 15 kb inserts). The In-Fusion Enzyme premix fuses PCR-generated sequences and linearized vectors efficiently and precisely, utilizing a 15 bp overlap at their ends. This 15 bp overlap can be engineered by designing custom primers for amplification of the desired sequences. This method can be used to clone single or multiple fragments into a single vector without sub-cloning.
In-Fusion HD Plus is a complete cloning solution that includes all the reagents required for successful cloning including the fast and high fidelity CloneAmp HiFi PCR Premix, the ultra-high efficiency Stellar competent cells, and a choice of PCR clean-up or gel extraction kit for DNA purification, along with the In-Fusion HD enzyme premix and reagents for control experiments as standard.
Features:
- Sub-cloning is unnecessary—Clone any insert, into any location, within any vector
- 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
Applications:
- PCR cloning
- HTP cloning
- Multiple fragment cloning
- Gene synthesis
- Gene design
- Mutagenesis
- Domain swapping
- Domain modification
In-Fusion® Cloning—a Fast, Efficient Way to Create Hundreds of Genetically-Encoded Biosensor Prototypes for Screening in Living Cells
Genetically-encoded, fluorescent biosensors have made it possible to look at signaling events in living cells. The greatest difficulty in creating FRET-based biosensors is finding the right way to fuse the donor and acceptor fluorophores to signaling proteins, or protein domains, without disrupting protein or cellular function. This application note describes the In-Fusion strategy, making it possible for a small team to create hundreds of biosensor prototypes quickly and efficiently.
Design Genes with Ease using In-Fusion® Cloning by Clontech Laboratories, a Takara Bio Company
In-Fusion can join any two pieces of DNA that contain a 15 bp overlap at their ends. The result is equivalent to a recombination event across the 15 bp overlap. The 15 bp overlap can be engineered by inclusion in forward and reverse primers used to PCR-amplify two segments of DNA. Originally described for inserting one piece of DNA into a restriction enzyme-digested plasmid, it has been found that In-Fusion can efficiently join up to four pieces of DNA in a single reaction. This application note explores the unique end-joining capabilities of In-Fusion to combine four fragments of DNA in a single, one-step reaction to develop seamless fusion proteins and modular expression vectors.
Cloning Blunt-End Fragments of Various Sizes
Takara’s Mighty Cloning Reagent Set (Blunt End) is designed to allow PCR products to be cloned into blunt-end vectors simply and quickly. Since PCR products with or without dA termini can be blunt-ended and phosphorylated at the 5’-ends in a single reaction, DNA fragments can be ready for ligation in a simple step. Pretreatment of PCR products to inactivate enzymes or remove unused dNTPs and primers is unnecessary. The cloning efficiency of the kit was demonstrated with PCR products that were used either directly or after gel purification.
High-Throughput Cloning of Functional Elements in the Human Genome
In this application note the In-Fusion® Dry-Down PCR Cloning Kit was used to insert a large number of putative promoter-containing DNA fragments from the human genome into a luciferase reporter vector for high-throughput screening of promoter activity.
Using In-Fusion® Technology for High-Throughput Precision Cloning of Constructs Expressing Signaling Pathway Protein Domains
In this application note the Oxford Protein Production Facility has created the pOPIN vector suite for use with Clontech’s In-Fusion cloning system.
