Pfu/Taq in PCR reactions

Duncan Clark Duncan at genesys.demon.co.uk
Mon Mar 6 05:45:18 EST 1995


Hang on Folks. The reason people did TA cloning was because Taq PCR 
products are very difficult to clone blunt because of the single base 
overhang. The single base was preferentially an A although the other 
bases are used. All the proof-reading enzymes do not appear to 
possess the terminal transferase activity so the PCR products can be 
cloned blunt without too much hassle. Now a Pfu/Taq mixture should 
give better fidelity but I would expect it to still add the single 
base overhang. For 2.5unit Taq there is only 1/64 unit Pfu so the 
terminal transferase activity should predominate.

At the end of the day I can still use a TA kit for cloning products 
amplified with Taq/Pfu mixes.

Duncan

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