RT-QuIC reactions were performed as previously described using recombinant bank vole PrPsen as substrate (residues 23 to 230; Methionine at residue 109; accession no. AF367624) . PrP from bank voles has proven to be a very good at detecting PrP amyloid seeding activity by RT-QuIC assay in a wide variety of species . Briefly, sample brains were homogenized to 10% (w/v) in PBS. Homogenate supernatants were then collected following a 2 min clearance step at 2000 x g. Samples were then 10-fold serially diluted in 0.05% SDS (sodium dodecyl sulfate, Sigma)/PBS/N2 (Gibco) to yield 10− 3 brain tissue concentrations. Four independent wells were tested for each mouse brain sample. 2 μl sample volumes were added to reaction wells of a black 96-well, clear bottom plate (Nunc) containing 98 μl of RT-QuIC reaction mix, resulting in final concentrations of 0.001% SDS, 10 mM phosphate buffer (pH 7.4), 300 mM NaCl, 0.1 mg/ml rPrPSen, 10 μM thioflavin T (ThT), 1 mM ethylenediaminetetraacetic acid tetrasodium salt (EDTA). The plate was then sealed with a plate sealer film (Nunc) and incubated at 42 °C in a BMG FLUOstar Omega plate reader with a repeating protocol of 1 min shaking (700 rpm double orbital) and 1 min rest throughout the indicated incubation time. ThT fluorescence measurements (450 +/− 10 nm excitation and 480 +/− 10 nm emission; bottom read) were taken every 45 min. Data was normalized to the percentage of maximum fluorescence obtained from the strongest signal in the run.