Abstract
A PCR cloning method that combines a dual selection pGATE-1 plasmid vector and an improved overlap extension cloningwasdeveloped. This efficient andcost-effective method allows for the introduction of DNA fragments into the Gateway cloning pipeline. The cloning efficiency is facilitated by a dual selection that includes the ccdB gene and gentamicin resistance. For users of the Gateway cloning system, substantial cost saving comes from eliminating BP recombination and ligation reactions to introduce DNA fragments into pDONR or pENTR vectors. Beyond the Gateway technology, this recombination-based cloning system can be used to efficiently clone PCR amplicons by adding 24-basepair adaptor sequences that are utilized by bacterial homologous recombination mechanism.
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