Patricia Anne Spears

Simultaneous SDA and Fluorescence Polarization Detection

5’-Fluorescently labelled, single-stranded oligodeoxynucleotides exhibit lower fluorescence polarization compared with their double-stranded counterparts because the fluorophore tumbles faster in solution when attached to single-stranded DNA. Consequently, fluorescence polarization is a convenient method of monitoring hybridization of detector probes without having to physically separate the single-and double-stranded forms. We have combined polarization detection with Strand Displacement Amplification (SDA) for real-time detection of target DNA sequences in a closed, homogeneous format.