Normalization In qPCR assay

Question

[Maria17]

Hi All,

Currently, I'm working on a qPCR assay.I normalized the targets according the housekeeping gene Ct mean and Log(2)Ratio for one target is 8 and other one is 5.I have some question Regarding the normalization:

1: is the higher log ratio is good or the lower one for the same gene in two targets?

2: can i calculate the log(10) ratio and what is the difference ?

3.i normalized the Ct mean, is it correct?

Normalization calculation:

1: x= Ct mean( House keeping gene - target)

2: x1=2^x

3: x2=x1*100

4: Ratio= log(x2,2)

Any recommendations or solutions would be appreciated.

Thanks

Maria

Answer 1

[Matthias Zepper]

Applied Biosystems always had a nice brochure on the basic theory of qPCR available - see Section VII for an excellent and comprehensive introduction. Also this paper might be helpful.

[Maria17]

Thank you.

[Matthias Zepper]

You're welcome.