How It Works

The In-Fusion Enzyme fuses PCR-generated sequences and linearized vectors efficiently and precisely by recognizing a 15 bp overlap at their ends. This 15 bp overlap can be engineered by designing custom primers for amplification of the desired sequences. Using this method, you can clone multiple fragments into a single vector without subcloning, create modular expression vectors with interchangeable parts, construct seamless fusion proteins, delete and replace DNA segments, insert point mutations, make internal fluorescent protein fusions, swap tags on a gene, add UTRs to a cDNA, insert restriction sites, and more.

Clone Into Any Vector

Clone directly into any vector at any site of linearization. Vectors can be linearized via inverse PCR or restriction digestion. If you do not already have a vector you would like to use, Clontech offers several In-Fusion Ready Vectors with options for fluorescent protein fusions, 6xHN tags for purification of recombinant proteins, and easy switching between expression systems.

Superior Cloning Efficiency

The In-Fusion system delivers greater than 90% cloning efficiency over a broad range of fragment sizes regardless of the nature of the DNA ends (sticky or blunt).

Multiple Fragment & HTP Cloning

With In-Fusion, multiple insert cloning is accomplished just as easily as single insert cloning. You can successfully combine not only two, but up to eight fragments of DNA in a single, one-step reaction (1, 2). The ability to rapidly and precisely clone in this manner makes the system highly amenable to automation. In-Fusion has been effectively applied in various high-throughput cloning projects, including work at Harvard Medical School (3), Stanford University School of Medicine (4), and the University of Oxford (5).