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.

In-Fusion Snap Assembly bundles components

All In-Fusion Snap Assembly products contain the In-Fusion Snap Assembly Master Mix and reagents for control experiments. The master mix is provided in a liquid format as a 5X enzyme premix in 20-μl to 100-μl aliquots, depending on kit size. The bundles also include:

• PrimeSTAR Max DNA Polymerase

  This convenient 2X liquid master mix offers exceptionally accurate, efficient, and fast DNA amplification. The premix contains dNTPs and an optimized buffer, allows rapid setup of PCR reactions, and facilitates successful cloning. The polymerase master mix is provided in 625-μl aliquots.

• Stellar Competent Cells

  High-efficiency competent cells are essential to the success of In-Fusion Cloning. ThisE. coli HST08 strain provides high transformation efficiency (greater than 5 x 10⁸ cfu/µg) and complements the efficiency of the In-Fusion Snap Assembly Master Mixes. Cells are provided in 100-μl aliquots in individual tubes.

• NucleoSpin Gel and PCR Clean-up kit

  This kit is suitable for gel extraction as well as PCR purification. Provided as individual columns.