14.8 Gateway® Cloning

Geneious Prime contains three operations to assist with Gateway® cloning. Gateway is a registered trademark of Invitrogen Corporation. The Gateway option under the Cloning menu will perform a BP reaction and/or an LR reaction on the selected documents. If there are a mixture of AttB/AttP and AttL/AttR sites on the input documents, it will perform a BP reaction on all documents with AttB/AttP sites, followed by an LR reaction on the results of the BP reaction and any input documents with AttL/AttR sites.

For example, to insert a PCR product with attB sites directly into a destination vector, select the PCR product, a donor vector, and a destination vector. Geneious will first produce an entry clone from the PCR product and donor vector, then react this entry clone with the destination vector to produce an expression clone.

    Annotate att sites...
    Add AttB Sites to PCR product
Annotate att sites...

This operation searches for AttB, AttP, AttL and AttR sites and annotates them on your sequence.

Add AttB Sites to PCR product

This operation allows you to add AttB sites to a PCR product. It will work on the following types of document: