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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