StarGate® Standard Cloning Principle in Detail

Step1: Standard Entry Cloning Step for Donor Vector generation.

The gene of interest (GOI) has to be equipped in a first PCR-step at both termini with combinatorial sites and the StarCombinase1™ recognition area which are important for oriented insertion of the PCR fragment into pENTRY-IBA51.
Recombination of the PCR product and the Entry Vector at the combinatorial sites (red and orange) leads to generation of the Donor Vector under loss of all StarCombinase1™ binding areas (dark orange with arrowheads) making the recombination reaction unidirectional and thereby highly efficient.

In the resulting Donor Vector the same combinatorial sites are now under control of StarCombinase2™ (light blue) thereby enabling a highly efficient and specific StarGate gene transfer process into Acceptor Vectors in a similar manner (Step2).

NEW Combi Entry Cloning Set (for appropriate primers use StarPrimer D´Signer Software) **

**Please note IBA has changed the Entry Cloning Step by May 2010. If you have used StarGate® in the past please check the sequence of your Entry Cloning Primers

References

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