what kind of quantification should I use? - (Jun/14/2007 )
I'd like to do a real time PCR to see if my gene of interest is expressed in several cell lines and in what quantity. Since I don't have "untreated" and "treated" samples, could I use absolute quantification? And also can I use plasmid with cDNA of that gene incorporated for standard curve and should I linearize it before. Also if I use absolute quantification, how do I normalize Ct of the gene of interest to a housekeeping gene?
You can also do relative quantification. You normalize against your housekeeping-gene (delta Ct) and directly compare the delta CT values between the different cell lines.