FIGURE 6.

Detection of capped transcripts abnormally accumulated in the lsm5/sad1 mutant. RACE-PCR analysis of AGI-code genes (A), intergenic non-AGI TUs (B), and antisense type of non-AGI TUs (C). Capped transcripts were examined by RNA ligase-mediated RACE. Total RNA was incubated with calf intestinal phosphatase to remove the 5′ phosphates, and an RNA adapter was ligated to the 5′ end of the transcript after a decapping reaction by tobacco acid pyrophosphatase (TAP) treatment. (+) and (-) indicate with and without TAP treatments, respectively. After the RT-reaction, using oligo(dT) primers, the first PCR was performed using an RNA adapter and transcript-specific primer sets, and then nested PCR was performed using a nested RNA adapter and first transcript-specific primer sets. TAP (-) indicates a negative control for RACE-PCR. Total RNA was extracted from heat-treated samples.