In-Fusion Snap Assembly products enable directional, seamless cloning of any PCR fragment—or multiple fragments—into any linearized vector in a single 15-minute reaction. No additional treatment of the PCR fragment (such as restriction digestion, ligation, phosphorylation, or blunt-end polishing) is required. The efficiency of In-Fusion Snap Assembly is over 95%, providing confidence in your clones and enabling you to scale up for high-throughput workflows.

The In-Fusion Snap Assembly Master Mix fuses PCR-generated sequences and linearized vectors efficiently and precisely—without the use of ligase—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 using our primer design tool. The In-Fusion master mix removes nucleotides from the 3" end of the linear DNA strands. This allows complementary base pairs between two pieces of DNA to anneal, leading to fragment joining. This method can be used to clone single or multiple fragments into a single vector without subcloning. Additional applications include mutagenesis, gene synthesis, gene design, domain swapping, and domain modification. The ability to clone directly into any destination vector at any locus (without the use of restriction enzymes) eliminates the need for any further subcloning or manipulation, simplifying the cloning workflow and further enabling a transition to high-throughput applications.

The In-Fusion Snap Assembly Master Mix is provided in a liquid format as a 5X enzyme premix in 20-μl to 500-μl aliquots, depending on kit size. It also contains reagents for control experiments.

Note: the 500-reaction and 1,000-reaction In-Fusion Snap Assembly Master Mixes contain two or four vials of the 250-reaction size, respectively.