Overview
Measurements for relative quantitative RT-PCR must be taken during the exponential phase of the reaction, when amplification is at its most efficient. In the first part of this protocol, the linear range of amplification is determined by carrying out 10 identical PCRs in the presence of [{alpha}-32P]dCTP and stopping one reaction after every two cycles. Amplification products are quantified on a denaturing polyacrylamide gel and the results plotted on a graph (counts per minute vs. cycle number). By Renée M. Horner.
Details
Details of this protocol, Relative RT-PCR: Determining The Linear Range Of Amplification And Optimizing The Primers:Competimers Ratio (Subscription Required), are located on a web site other than Biocompare Protocols.