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.