6/19/2002 Juli Grose
PfuTurbo DNA Polymerase (Stratagene catalog #600250) is an "enhanced" version of Pfu DNA polymerase for high-fidelity PCR (for details see the Stratagene instruction manual). Pfu has significantly lower error rates (in part due to its proofreading ability).
Use Pfu as described below and only if NECESSARY (ie when cloning). Efficient sequencing may be done by running 100 uL reaction with TAQ in 5 separate capillary tubes (thus reducing the probability of an abundant product with a bp change due to PCR).
Keep Pfu on ice...it is EXPENSIVE.
1. Standard reaction mixture (scale down for trial reactions)
2. Standard PCR cycling program (Idaho Tech AirCycler)
Note: The fifteen second holds during cycling are essential for efficient PCR
Any Questions? Ask Juli!