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What type of cloning strategy should I use with a fragment generated using PCRBIO HS VeriFi™ polymerase?

PCR products generated by VeriFi™ Hot Start Polymerase have blunt ends, making them suitable for blunt end cloning without requiring any further modification.

If you would like to add A-overhangs, you can use Taq DNA Polymerase or Klenow (exo-) DNA followed by TA cloning1. Before proceeding with the addition of A-overhangs, ensure the VeriFi™ Hot Start Polymerase has been removed from the reaction. If there is any VeriFi™ Hot Start Polymerase present, the 3’-5’ exonuclease activity of the enzyme can remove A-overhangs.

1  Yao, S., Hart, D. J. & An, Y. Recent advances in universal TA cloning methods for use in function studies. Protein Eng Des Sel, doi:10.1093/protein/gzw047 (2016).

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