Takara Site-directed mutagenesis

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.

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Overview

• A single system for deletions, base substitutions, or additions
• Flexible enough to use with any vector
• Over 95% accuracy, guaranteed

More Information

Please see the product`s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.