I am still learning the nitty-gritty of performing a RTPCR...I usually run qRTPCR by the Taqman method, comparing gene expression levels between GFP and siRNA treated cells and look for downregulation. I use
RNAse P as the internal control for normalization and use transfected HEY cell line.
However, I m now confused as I am required to check for my gene's expression level in 5 different
cancer cell lines. Since i just have to quantitate the expression and not really compare against anything,
I am a little lost about how shd I go about running the samples and calculating using excel sheet.
SHould i consider one of the cell line as baseline i.e 1 and check expression levels?
and what shd be the other controls?
not sure if I am putting across my question effectively...can anyone reply please?
Thanks for any help...