Gibson assembly enables seamless joining of multiple DNA fragments using a 5′ exonuclease, DNA polymerase, and DNA ligase in a single isothermal reaction. Fragments must share ≥20 bp homology with adjacent segments (Tm ≥50°C).
Principle: Arrange DNA fragments in desired assembly order. Including vector sequence at both ends streamlines circular construct design.
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Primer Design Options
Need details? Sequence markup, parameters, output columns and the suggested workflow are documented in Help: Gibson Assembly tool · Help: Troubleshooting