Fig. 3

Characterization of the spliced product using S1 nuclease. a Schematic representation of S1 nuclease protection assay. Top, a diagram of the hybrid formed between spliced RNA and DNA oligonucleotide (50 nt) complementary to exons junction. Bottom, A diagram of protected sequences after S1 nuclease digestion. b Spliced [³²P]-RNA produced in in vitro splicing assay was gel purified as described in Materials and methods. [³²P]-pre-mRNA (negative control) and spliced product [³²P]-RNA were hybridized to oligo DNA that is complementary to exons junction. Following hybridization, [³²P]-RNA–DNA hybrids were digested with S1 nuclease that degraded single stranded nucleic acids. The size of the protected region (50 nts) is indicated. Red arrowhead shows protected exon junction sequence in the spliced product