Gibson Assembly in Geneious Prime

Perform Gibson Assembly

Prepare multiple DNA fragments, linearize your vector at the desired site, simulate Gibson assembly, and perform batch cloning from a sequence list.

LENGTH 7 minutes

Common Questions about Gibson Assembly

What does Geneious Prime simulate in a Gibson Assembly reaction?

Geneious Prime simulates the full Gibson Assembly workflow: linearizing the vector at the insertion site, preparing insert fragments, configuring exonuclease type and overlap parameters, generating primers with homologous extensions, and producing a Gibson Report that validates overlap regions, melting temperatures, and scarless joint formation. The simulation confirms fragment assembly before any bench work begins.

Does Gibson Assembly in Geneious Prime also support related isothermal methods?

Geneious Prime's Gibson Assembly tool also supports related homology-based cloning methods including SLIC, CPEC, In-Fusion, and other isothermal approaches that share the same overlapping-fragment assembly principle. Researchers working across multiple ligation-independent cloning strategies can use the same interface for each method.

Does Geneious Prime check that Gibson Assembly overlaps are long enough?

Geneious Prime validates that complementary overlap regions between fragments meet the recommended minimum of 18 bp, calculates melting temperatures for primer binding sites and extensions, and reports on primer characteristics and binding sequences. The Gibson Report flags overlap issues before you proceed, reducing the risk of failed assemblies caused by insufficient homology.

How does Geneious Prime generate Gibson Assembly primers?

Geneious Prime automatically designs PCR primers with the homologous extensions required for Gibson Assembly. The tool generates "Gibson Primer Extension" annotations for any custom sequence additions and calculates Tm for both the binding region and the extension, ensuring primers are compatible with the isothermal assembly conditions before synthesis.

Can I perform batch Gibson Assembly in Geneious Prime?

Geneious Prime supports batch Gibson Assembly using sequence lists, allowing you to insert multiple insert variants into the same vector position in a single operation. This is useful for testing libraries of inserts or validating multiple constructs in parallel without repeating the setup for each variant individually.

Can Geneious Prime output ready-to-order inserts instead of PCR primers for Gibson Assembly?

Yes. Switching from "design primers" to "synthesize inserts" in the Gibson Assembly options skips PCR primer design and instead extends the required homologous overlaps directly onto the insert sequence, producing a construct that can be submitted straight to a gene synthesis vendor rather than built by PCR.