Can I clone multiple fragments into one vector in a single In-Fusion Cloning reaction?
Can I clone multiple fragments into one vector in a single In-Fusion Cloning reaction?
Yes—we have successfully tested multiple-fragment cloning with up to five inserts (see the figure and table below for cloning schematic and colony screen results, respectively).
Primer design for multiple-fragment cloning can be done with our online Primer Design Tool. (The Primer Design Tool is compatible with Mozilla Firefox or Google Chrome web browsers, but not with Internet Explorer.) Please note that between two adjacent fragments, only one homologous overlap is required for the In-Fusion reaction. This overlap can be located on either of the fragments.
In-Fusion Cloning has an improved capability for cloning multiple fragments in a single reaction. Using this system, cloning up to four 1-kb fragments simultaneously is as easy as cloning a single fragment. This saves weeks that would otherwise be spent screening clones and subcloning.