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In-Fusion Cloning¶

What is In-Fusion Cloning?¶

In-Fusion is an overlap-based assembly method that joins DNA fragments sharing short terminal homologies of at least 15bp. The principle is similar to Gibson Assembly:

  • A 3'→5' exonuclease exposes single-stranded overhangs at the ends of the fragments
  • Complementary overhangs anneal
  • The product is transformed into bacteria that anneal and repair the fragments, producing a circular plasmid.

How to plan In-Fusion using OpenCloning¶

Like any other cloning method, click on the plus icon below a sequence in the Cloning tab and select In-Fusion Cloning. Then, select the sequences to combine in the Assembly inputs field. OpenCloning evaluates adjacent fragments for sufficient homologous overlaps in the intended orientation and proposes valid assemblies.

If you need to add overlaps by PCR before assembly, see Primer design for Gibson assembly. The same style of terminal overlaps is used for In‑Fusion.