Fig. 1. Comparison of the padlock probe-based isothermal amplification assay with the RT-PCR assay for detection of the two-step RNA splicing products. The major difference is that the amplified assay can be used for simultaneous detection of the intermediate and final products, while RT-PCR could not detect the intermediate products. (a) Fluorescence responses of the isothermal amplified assay under different conditions: in vitro splicing assays were conducted in the presence of NE (red curve) or absence of NE (black curve). The splicing products were analysed by the padlock probe-based isothermal amplified assay. (b) Gel electrophoresis of the RT-PCR products for in vitro RNA splicing: 40 pM CDC pre-mRNA was incubated in the presence of NE (lane 1) or absence of NE (lane 2) for 1 h at 30 °C under the conditions of in vitro RNA splicing; lane 3, spliced mRNA; lane 4, NE.
