Interpreting QPCR DATA

[George25]

Hi to all,

I am new in QPCR and I have some questions.. Last week I did my first QPCR using TaqMan probes. I did relative quantification and so I didnt do a standard curve.. So for quantification I try to use the ΔΔCt method (comparative CT method). The real time PCR report gives me the ΔΔCt values for each of my sample and then I calculate the 2-ΔΔCT that can be used for my graph. Although, when I do the ΔCt and ΔΔCt calculations according to the manifacturer's instructions by hand I have different 2-ΔΔCT values and hence different expression profiles for my samples. Could someone gime me advice on that?
Also, what other methods do you use for relative quantification? Which one better in my case.

Thank you very much

[Aga]

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