PCR quantification - (Jan/28/2009 )
Hello all wise peoples. Im conducting end point PCRs using a number of primers pairs which im comparing to a baseline reading constant between conditions. Im told that using image J and some of its associated plugins i can quantify my pcr bands and and produce ratios relative to the baseline which can then be compared to each other. I had a search on the web but didnt realy manage to find any real instructions. Can anyone help, either by explaining the process itself or pointing me in the direction of someone/something that can? Any assistance would be realy appreciated, quantifying my results would add much weight to the project im doing.
Sorry of not being of direct help, but have you considered using real time PCR, if a machine is available to you? Quantitation is much more accurate & reliable using rtPCR data than band intensity data (even if it is digitised).