Cells were transfected with the minicassette plasmid SXN13 (23 (link)) with Effectene (Qiagen). After 48 h, total RNA was extracted with miRNeasy columns (Qiagen) and cDNA prepared using oligo-dT and Superscript III (Life Technologies). PCR reactions to detect the splicing products were done using the primers SNXF: 5′-GACCATTCACCACATTGGTG-3′; and SXNRv: 5′-GAACCTCTGGGTCCAAGG-3′. PCR for specifically detecting partial splice products used SXNF and SXNJrv: 5′-GACCACCAGCAGCCTGGA-3′ or SKNJF: 5′-GCCCTGGGCAGGTCGAC-3′ and SXNRv. All products were electrophoresed in a 2% agarose gel.
PCR Primers (Table1 ):
PCR Primers (Table
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