What PCR polymerases are recommended for amplification of the In-Fusion cloning insert?
What PCR polymerases are recommended for amplification of the In-Fusion cloning insert?
In-Fusion Cloning is compatible with any PCR polymerase. 3' overhangs do not interfere with the cloning reaction.
To ensure an error-free insert, use a polymerase with high proofreading activity, like PrimeSTAR Max DNA Polymerase (supplied with some In?Fusion Snap Assembly bundles). This polymerase is highly robust and accurate, enabling amplification of up to 6 kb of human genomic DNA, 10 kb of E. coli genomic DNA, and 15 kb of lambda DNA. It is compatible with two- or three-step PCR cycling, and exhibits minimal error rates on GC-rich templates.