Question

how to enhance the amplifiaction of pcr product in real time pcr

Answer 1

I'd love to have that much DNA! I frequently amplify from template which has undetectable levels of nucleic acid. Have you looked at your sample on a nanodrop? You may have contaminants which are inhibiting your PCR- in which case, diluting your template may help. I know it sounds wrong, but diluting the sample will dilute the contaminants and you'll still have enough DNA. If I suspect I have PCR inhibitors in my sample, I dilute it 1:1, 1:10 and 1:100. Otherwise, optimise your PCR reaction as Guarav has suggested. If I'm really struggling then I'll try different kinds of Taq, or just loads and loads of taq (Not recommended, but a desperate last measure).

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