Figure 3.

Determination of the exact hybridization sites of random oligonucleotides in the nucleotide 170-350 region by radiolabeled primer PCR analysis (see Materials and Methods). The RNA template (561 nucleotides long) was subjected to RT-ROL, then the exact sites of hybridization were determined by amplifying the RT-ROL products with radiolabeled primer sROD145 and unlabeled TAG oligonucleotide. Lanes C,U,G are sequencing lanes; Lane 1: RT-ROL reaction without any added oligonucleotides; Lane 2: positive control reaction in which an oligonucleotide complementary to the PBS region was added to the reverse transcription reaction instead of the random oligonucleotides; Lane 3: complete RT-ROL reaction showing several amplification products arising from binding of random oligonucleotides and subsequent reverse transcription and PCR amplification.