t-test & qRT-PCR - (Feb/05/2015 )

I've been reading a lot of how to analyse qRT-PCR results. So far I have calculated:

-Delta Delta Ct - therefore know whether my target gene is up/down-regulated

I was looking at other documents online which split the target and controls up by repeats in a table and calculate everything separately (compared to the method of finding the mean of the Ct values and working through it). I wish to perform the t-test on my data but am not sure how to do so as some sources say that you:

1) Write your experiments are separate values

2) Subsequently calculate means

3) Average delta delta Ct values & perform t-test on those

An example of my trouble in reference to the attached image is:

1) When I calculated the Delta Delta Ct value for the whole experiment, I got the same answer as the Target treatment which is 0.046, but no idea how to get 1.002 and even their ΔΔCq
Expression for Non-Targeting Controls do not match: (2^-0.1 is different - 0.960 & 0.945??)

My main aim is from my data to be able to:

1) Calculate delta delta Ct value (Done)

2) Standard Deviation - can do if I do everything separately

3) t-test

If anyone has any help for steps two and three - would appreciate it. thanks.

Double-post by accident, apologies.

merged them

hobglobin