This is a stupid but quite technical question.
We have a PCR machine that has 20 uL minimum reaction, so it can't be set into 15 or 10 uL in the machine.
Is this going to affect the result ?
Thank you very much
It should not. Reaction volume might influence the time required for the solution to reach the set temperature, thus affecting the duration of PCR. I expect, that setting 20 µl and physically putting 100 µl into the tube might not be the best idea. But the other way, this setting should not be important.
I agree with Ruta Gerasimaite it should not make much difference when your volume is less than the machine set volume. Most pcr machines assume a volume inertia so overshoot the set temperatures knowing the sample temperature lags behind the temperature profile so from 94 to annealing your sample may drop slightly below the set annealing temperature but probably not by more than 1c and most pcr reactions are not that sensitive so expect it to work and if there is any non specific amplification adjust the annealing temperature up a little
Thanks for the answers Ruta Gerasimaite and Paul Rutland . It's very clear. I will try to do few samples first.
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