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Nested PCR - no amplicon - (Feb/15/2006 )


I´ve made a nested PCR but can´t get an Amplicon after the 2. Step.
Anybody can solve me with this problem?

Thanks, Greets



If you don't obtain an amplicon in your 2nd PCR maybe your primers don't work. How did you design them? Did you blast them against your 1st amplicon after the design?

Do you migrate in agarose your first PCR? Do you see your amplicon? How intense is it? 2 thing s may happen: too much or little DNA added for your 2nd PCR.

Lots of things may happen...