Perform solid-phase capture and sequence-specific detection of PCR products in an easy-to-use, automatable format and allele-specific hybridization for the detection and genotyping of point mutations. PCR amplification is carried out with one 5′-biotinylated primer and one unlabeled primer. The amplified products are immobilized in streptavidin-coated strip-well plates. The non-biotinylated strand is removed by sodium hydroxide denaturation, and the biotinylated strand is hybridized to a sequence-specific fluorescein-labeled probe. After a wash step, the probe is detected with a peroxidase-conjugated anti-fluorescein antibody and the chromogenic peroxidase substrate TMB. The assay can be completed in 2.5 hours.
Purchase of this product is accompanied by a limited license for use in the Polymerase Chain Reaction (PCR) process for research purposes only and in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by an up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., and authorized thermal cycler.