I have been doing RT-PCR , and in I use standard curve method to quantify. in every PCR plate I run, i was taught that I should run corresponding REFERENCE GENE triplicates. Just not for any given cDNA synthesis, but for every PCR run I included a triplicate of 18S.
However i was recently told that this is a common misconception and that I do not need to do this for every plate, but only for a given cDNA synthesis, and then I can use these reference gene values to normalise genes of interest values.
I would be very gtrateful if any one who is using STANDARD CURVE METHOD COMMENT ON THIS.
What are common sources of contamination in the qRTPCR? I too would be grateful for any feedback to this dilemma? thanks
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