PCR Cycle

Vellanoweth's Lab ddeloss at calstatela.edu
Fri Mar 19 15:50:33 EST 1999


I am trying to amplify a gene that is 3000bp long using genomic plant
DNA and I would like to know what the best times for each step in the
cycle would be.  The annealing temperature of the primers is 57oC and
the current cycle I use is the following:
 1.  95oC - 1 min
 2.  40oC - 4 min
 3.  72oC - 1 min
 4.  95oC - 45 sec
 5.  57oC - 2 min
 6.  72oC - 1 min
Then 34 cycles to step 4
 I was told that perhaps a touchdown PCR would be better but I also
don't know the times for each temperature for that procedure.  You can
reach me here or through email at <cgama at calstatela.edu>.  Any help
would be greatly appreciated.
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