Figure 3.

Quantification of the RT-PCR products by PCR kinetic analysis cDNAs were obtained from total adult rat liver RNA as described in Materials and Methods. PCR amplifications were performed with E1f and E5r primers (Table 1) in the presence of 2.5 μCi (0.25 μl) of [α-³²P]dCTP (300 Ci/mmol). After 20 amplification cycles, a small portion of the PCR mix (10%), was removed every two cycles and the products were resolved on 8% acrylamide gels. The amount of radioactivity in each band was determined by scintillation counting. For kinetic analysis, values of log counts per minute were plotted against the number of cycles. The antilog of the respective intercepts shows the proportion of each mRNA. The upper straight line denotes the 523 pb mRNA (○), and the lower lines correspond to the 659 bp (□) and 784 bp mRNA (▵). The autoradiography is shown in the Inset.