Takara In-Fusion Snap Assembly Master Mix \10 Rxns

In-Fusion Snap Assembly Master Mix is designed for fast, directional cloning of one or more fragments of DNA into any vector. This proprietary master mix fuses DNA fragments (e.g., 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 into the primers designed for PCR amplification of the desired sequences. In Fusion Snap Assembly Master Mix offers high efficiency, even for applications that can be challenging, including the cloning of long fragments, short oligonucleotides, and multiple fragments.

Performance comparison between In-Fusion Snap Assembly and NEBuilder HiFi using inverse PCR.A single 3.8-kb insert (Panel A) or a 34.2-kb adenovirus insert (Panel B) was cloned into a 2.7-kb vector which was linearized via inverse PCR. These cloning reactions were performed in triplicate with both In-Fusion Snap Assembly and NEBuilder HiFi. Primers were designed according to the manufacturers specifications. After transformation and plating, 20 colonies from each replicate were analyzed by Sanger sequencing (for the 3.8-kb insert) or colony PCR (for the adenovirus insert) to determine the cloning accuracy. In-Fusion Snap Assembly yielded 2X more colonies than NEBuilder HiFi.

The In-Fusion cloning protocol.