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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