In-Fusion Cloning products provide the flexibility to perform site-directed mutagenesis (deletions, base substitutions, or additions), in addition to powering any of your single- and multiple-insert cloning experiments. In-Fusion Cloning makes it easy to perform mutagenesis by combining the power of In-Fusion technology with inverse PCR, a method for rapid in vitro amplification of the DNA sequences that flank a region with a known sequence.
In-Fusion Cloning products provide the flexibility to perform site-directed mutagenesis (deletions, base substitutions, or additions), in addition to powering any of your single- and multiple-insert cloning experiments. In-Fusion Cloning makes it easy to perform mutagenesis by combining the power of In-Fusion technology with inverse PCR, a method for rapid in vitro amplification of the DNA sequences that flank a region with a known sequence.
During inverse PCR, primers are oriented in opposite directions on your circular cloning vector, so to perform mutagenesis with In-Fusion systems, design your PCR primers so that they have a 15-bp overlap with each other at their 5" ends and incorporate the mutation of interest. Use the PrimeSTAR Max DNA Polymerase (a high-fidelity PCR polymerase included with all In-Fusion Snap Assembly bundles) to perform your PCR, add the In-Fusion Snap Assembly Master Mix to your linearized PCR product, and transform into the provided Stellar Competent Cells. You have a ≥95% chance of recovering your final desired construct—the first time and every time.
