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FAQs
Browse our frequently asked technical questions by selecting a product below.
What type of cloning strategy should I use with a fragment generated using PCRBIO HiFi polymerase?
PCR products generated by PCRBIO HiFi 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 PCRBIO HiFi polymerase has been removed from the reaction. If there is any PCRBIO HiFi 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).
