Have you tried the 2^-delta delta Ct equation to monitor the fold change and then run a t test on the data?

Hi,

First of all, you must run an 8 point standard curve with the different primers you are using for your RealTime PCR. For this make serial dilutions in 4 or 10 steps depending on the representation of your gene of interest. With this standard curve, you will be able to calculate the Efficiency of your reaction. 

In the case of the 2^-delta CT is always the assumption, that your efficiency = 100% what is the same as saying that you will have after each cycle 2x more DNA. This efficiency is not always the truth.

After calculating your efficiency you will be able to calculate the relative expression for each of your samples Eff^-Ct. With this values, you can normalize to the gene of interest and your control in the same way you work with any other experiment.

For the complete system on RealTime OCR have a look to the link.

Greetings

Xiomara

https://www.ncbi.nlm.nih.gov/pubmed/19246619

Hi,

Have you checked this?

Article BootstRatio: A web-based statistical analysis of fold-change...