I am trying to design lamp PCR primers and I am really confused about their annealing temperatures. Annealing temp for the same region in PCR is 58 degrees C but in lamp PCR primer designing different primer temperatures are shown for different primers. Is that right or not?
hello
that’s possible to have same annealing temperature. anyway, you can check your primers using gradient PCR (with set up gradient annealing temperature). from your result yku can get your annealing temperature of your primer
My advice: Do not worry too much about theoretical Annealing Temperature (as long as they are not too different for both primers). It's quite common to get a different Tann for the same primer depending on the software/tool you use. You should ask for them, and once in your hands, find out the optimal amplification conditions through a gradient PCR.
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