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single tube nested pcr

gzk at ollamh.ucd.ie gzk at ollamh.ucd.ie
Tue Feb 18 11:40:42 EST 1997

I'm looking for some advice on setting  up a single-tube nested PCR assay.
The protocol I'm following has the first set of primers at a lower concentration
and higher annealing temperature.  The annealing temperatures of my primers
are higher in the second round.  Any idea if it is preferable to have the 
first annealing temperature higher or not.  Of course I could just try it
and see but any advice would be useful before I try.
Geraldine Kaminski

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