Generating Overhangs

If generating overhangs for Golden Gate Assembly outside of YTK/BTK framework, NEB's GETSET tool can be useful for generating a set of high fidelity overhangs to use. If compatibility with other overhangs is necessary required overhangs may be set.

For scarless assembly NEB's SPLITSET tool may be used. You'll need to input the DNA sequence you're assembling and specify the number of assembly fragments. The tool will find high fidelity overhangs that will allow you to scarlessly assembly your DNA using Golden Gate Assembly.

These tools can be useful for domesticating part plasmids, for creating high-fidelity subparts for an existing assembly schema (such as the BTK/YTK), and for creating scarless assemblies.

-- Victor Li - 2021-11-03
-- Updated by Alexa Morton - 2025-06-17