Figure 1.

Development of RT-PCR experimental conditions. A and B, Ubq10 is a constitutive control. Four-day-old Arabidopsis dark-grown wild-type seedlings were kept in the dark (Dark) or exposed to 1 h light (Light), RNA was extracted, and amounts of Ubq10 were determined by conventional RT-PCR followed by Southern blot (A) or by the LightCycler (B). The amount of Ubq10 PCR products as a function of polymerization cycles is shown in A. The calculated initial amount of Ubq10 transcript is shown in B. C and D, RT-PCR controls for Elip1 and Elip2. C, Elip1 and Elip2 primer pairs were used for PCR on genomic DNA and cDNA (from “Light” above) templates, and in the absence of RT (−RT). D, “Light” sample from above was used as template to determine the kinetics of PC reaction as a function of number of PCR cycles, as detected by dot blot.