RF-Cloning.org: an online tool for the design of restriction-free cloning projects

Restriction-free cloning (RF-cloning) is a PCR-based technology that expands on the QuikChange™ mutagenesis process originally popularized by Stratagene in the mid-1990s, and allows the insertion of essentially any sequence into any plasmid at any location. While RF-cloning is a powerful tool for the design of custom plasmids when restriction sites are not conveniently situated, manually designing the requisite primers can be tedious and error prone. We present here a web-service that automates the primer design process, along with a user interface that includes a number of useful tools for managing both the input sequences and the resulting outputs. RF-Cloning is free and open to all users, and can be accessed at http://www.rf-cloning.org.