PCR getting smear

K J C 944 kjc944 at aol.com
Sun Jun 16 20:37:35 EST 1996


Zahra,
You might also try touchdown PCR.  Run your reaction at several different
temperatures to determine the highest annealing temp at which you still
get a band.  Then do a touchdown reaction starting with an annealing temp
just above this upper limit, lowering the annealing temp by 0.5 degrees
each cycle until the optimal Tm of the primer is reached.  Minimize the
number of cycles at your lowest Tm to reduce the amount of nonspecific
products.
good luck!

Kathryn J. Coyne
College of Marine Studies
University of DE
Lewes, DE  19958  



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