I would like to know more about validating my semiquantitative RTPCR
system. From the same RT sample I am doing two PCR reactions, one
using primers for the message of interest and one for
B2-microglobulin, which should act as a control. The PCR reactions
are done under the same conditions, at the same time. I have
established that the cycle number has been limited to ensure that
amplification is still exponential.
Is it valid then to compare the intensity of staining of the ethidium
bromide stained bands and express results relative to the B2M control?
Any references on principles involved would be appreciated