Phage plaque PCR

Sasha Kraev bckraev at aeolus.ethz.ch
Thu Oct 13 05:08:30 EST 1994


Hi, Felix
There is no need for a special buffer, just suspend your plaque, preferably
from a fresh plate, in 50-200 ul sterile water, freeze-thaw once, mix by
tapping with a finger, and remove 2 ul into a pre-assembled 50 ul PCR reaction
without an enzyme. Start the program with a 5 min 95 C heating step, then pause
and add polymerase. Continue with your program as usual for 30 cycles. Can
also remove aliquots from an SM-eluted plaque, but that changes Mg 
concentration in the PCR and may lead to extra bands etc. Good luck, Sasha



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