Gateway Cloning
Course Content
Gateway Cloning with SnapGene
Gateway Cloning with Multiple Inserts
Common Questions About Using SnapGene
Which Gateway reactions can SnapGene simulate?
SnapGene can simulate BP reactions (creating Entry Clones from attB-flanked inserts and Donor Vectors), LR reactions (transferring inserts from Entry Clones to Destination Vectors), or BP + LR reactions in a single operation. For multiple inserts, SnapGene designs attB-linked PCR primers for each fragment and simulates the complete workflow through to an Expression Clone.
Does SnapGene include Gateway donor and destination vector sequences?
Yes. SnapGene provides a library of commonly used Gateway donor and destination vectors, accessible directly within the cloning dialog. A custom donor or destination vector from your own sequence files can also be selected if the standard library does not include the vector you need.
Can SnapGene design the attB-linked PCR primers needed for Gateway cloning?
Yes. During the BP reaction simulation, SnapGene designs appropriate PCR primers with attB sites added to each insert fragment. Primer names can be edited in the attB Fragments tab, and sequences can be exported for ordering.
Does SnapGene support multi-insert Gateway cloning?
SnapGene supports cloning with multiple insert fragments via BP + LR reactions in a single operation. However, it cannot currently simulate Gateway cloning with vectors containing more than one pair of att sites. The support documentation confirms this as a current limitation.
How does SnapGene display a multi-insert Gateway product?
In the Expression Clone tab, inserts are coloured alternately red and blue in Map view. Switching to Sequence view allows you to scroll through fusion boundaries between inserts and confirm reading frame continuity at each junction.
What is the difference between this simulation series and the Gateway Cloning topic guide?
The simulation series focuses on the software workflow: running BP and LR reactions in SnapGene, designing attB primers, and producing a verified product file. The Gateway Cloning topic guide explains the underlying technique: how att site recombination works, the roles of Entry and Destination vectors, and practical guidance for designing Gateway experiments.
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