FAQ: Can I PCR-amplify the assembled product?

Yes. The assembled DNA molecule is covalently joined and may be PCR-amplified. Additionally, if the final product is a closed circular DNA molecule, it may be used as a template in rolling-circle amplification (RCA).
  1. NEBTVToolsThumbnailNEBuilderAssemblyTool20Fragments Amplified by PCR

    NEBUILDER® Assembly Tool 2.0 Fragments Amplified by PCR

    This video demonstrates how the NEBuilder Assembly Tool can be used to generate overlap sequences for the assembly of two fragments into a vector. In this scenario, all fragments are amplified by PCR and the vector does not have convenient restriction sites.