RT-QuIC analysis of uninoculated and sCJD inoculated transgenic mice. All brain samples were tested at a 10−3 dilution. In A–F, four independent wells from each mouse are represented as individual curves with unique symbols. A–E Data from five different uninoculated tg66 and tgRM mice tested with Ha rPrP substrate. F An example of an uninoculated tg66 mouse tested with BV rPrP substrate. G Ha rPrP RT-QuIC data from several replicate trays of uninoculated transgenic mice (left side) and sCJD-positive control (right side) transgenic mice at the 25-h timepoint. Each point represents measured fluorescence in a single test well. Different tray numbers and different mice are indicated below the x-axis. A horizontal dashed line is shown at 10% florescence (27 standard deviations above baseline florescence) that clearly separated negative and positive reactions.