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. 2016 Sep 26;6:33920. doi: 10.1038/srep33920

Figure 2. The minigene splicing assay based on the pSPL3 exon trapping vector.

Figure 2

(A) The pSPL3 vector contains two exons SD and SA, and a functional intron, with transcription beginning following the SV40 promoter and ending at the LPAS (late poly (A) signal). Wild pSPL3-W and mutant pSPL3-M plasmids containing 207 bp of intron 7, 136 bp of exon 7 and 186 bp of intron 8 were separately cloned into the XhoI and NheI cloning sites of the pSPL3 vector. (B) Agarose gel electrophoresis of RT-PCR products. SD6 and SA2 primers were designed for RT-PCR amplification of cDNA sequences generated by transfected 293 T cells. Lane1: Marker; Lane2: empty vector (263 bp); Lane3: 399 bp (263 bp + 136 bp) and 263 bp; Lane 4: 263 bp. MCS: multiple cloning sites.