PCR Question

[Michael Cooley]

Jim Kyriannis (jim at phri.nyu.edu) wrote:

: I have 2 30-mer PCR primers which have a Tm higher than 100 degrees.  I
: have tried to use these two primers to PCR directly from yeast colonies with no
: success.  I have used other primers under the same conditions, which have
: worked just fine.  Could the high Tm be the source of the PCR reaction
: failures?


: Thank you.

: Jimmy

It is impossible to have a Tm of greater than 100 oC even if it were 100 
% GC.