REAL TIME PCR 7005 v 2.01 : I need more reference samples! - How to add more than one reference sample to Real Time?? (Aug/12/2009 )
I have a problem. We just instal the new version of Applied Biosystem Real Time PCR program (7500 v2.01) in the lab. And we want to use the quantitative, delta delta Ct program. However, our experiments demain more than one control, or reference sample (as named in the 'help' program), because most of our research include a time course experiment, wich means, for example, more than one point of plant development wich demaind a control and a treated sample. No one could understand how add more than ONE reference sample in the program. I understand that major experiments need just one reference, but we lead with plants and considers the circadianum cycle and the developmental differences between samples. We are just this close to understand that this program (based in the delta delta CT) do not allow this kind of alteration.
There's someone with this kind of knowleadge here?
I and all braziliam fellows will pray forever for the help!
my personal e-mail: firstname.lastname@example.org
The control you mentioned in your text is referring to the untreated sample in your experiment right?
The reference sample refers to those endogenous target or genes that are usually do not or have minimal change in their expression level with or without any treatment. This reference sample will serve as an input.
To any other researcher here, please correct me if I am wrong.