Dear Alaa

• I am the above person referred to in the previous answerr (so thank you Saurabh !!)
• You should absolutely use 2^ ddCt not 2^ct in which I assume you would be just be comparing the difference in Ct value between your WT and mutant mouse; This would not normalise against a housekeeper which you absolutely need to do when comparing 2 different cDNA sources (biological replicates)
• Even so, the 2^DDCt method is really for comparing the same model treated and intreated not strictly speaking 2 different systems (mice)
• You woiuld be better determining absolute expression by standard curve in the 2 different systems and then computing the quotient to obtain expression difference
• Take a look at the guide I have attached for absolute expression (using standard curves)

You can go through the answer of two people, they explain nicely in context of qPCR data analysis.

https://www.researchgate.net/profile/Laurence_Dawkins-Hall

Jochen Wilhelm

Dear Alaa

• I am the above person referred to in the previous answerr (so thank you Saurabh !!)
• You should absolutely use 2^ ddCt not 2^ct in which I assume you would be just be comparing the difference in Ct value between your WT and mutant mouse; This would not normalise against a housekeeper which you absolutely need to do when comparing 2 different cDNA sources (biological replicates)
• Even so, the 2^DDCt method is really for comparing the same model treated and intreated not strictly speaking 2 different systems (mice)
• You woiuld be better determining absolute expression by standard curve in the 2 different systems and then computing the quotient to obtain expression difference
• Take a look at the guide I have attached for absolute expression (using standard curves)

Dear Laurence!

Thanks a lot for your very very kind reply!! I appreciate it very much! I took a look at the PDF and am following it!

All the best to you! Thanks again

Alaa