ASPCR

Alternative splicing analysis by end-point RT-PCR, or ASPCR, involves the relative quantification of alternative isoforms from a RT-PCR reaction. We design primers to amplify both isoforms in the same reaction tube. The splicing event is thus quantified by capillary electrophoresis from the ratio of the longer isoform (inclusion) against the sum of the shorter (exclusion) and longer isoforms. This ratio is dubbed "percent splicing index", or PSI. PSI data is presented in bar graphs, heatmaps and data sheets.

  • Perkin Elmer LabChip GX Touch HT capilary electrophoresis system (three units).
  • Optimal detection and quantification for 80 to 650 bps amplicons with at least 15% size separation.
  • 2500 reactions daily throughput.

Applications

  • Alternative splicing event quantification analysis from selected events or from genes splicing survey.
  • RNA-Seq splicing validation by RT-PCR.
  • Bacterial molecular strain typing by ribotyping and multi-locus variable-number tandem repeat analysis (MLVA).