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How much of the starting amounts for Real time PCR? - (Apr/24/2005 )

After i have reversed transcribed my mRNA samples to cDNA.
How much of cDNA samples should i add to run real time PCR of both untreated and treated samples? If we add the same amounts of both types of samples then how can we see the difference of gene expression profiles? blink.gif

Thank you for any ideas.


Dear friend,

You are right. You will not see see the different. So you just go by volume, let say 2ul each samples.

I would like to suggest you to use one-step RT-PCR. The reason is, when you do RT step seperately from PCR, the dilution factor in the RT step has to be take into consideration when you set your standard.