This package contains installers for two programs that perform single-molecule kinetic fingerprinting (SiMREPS=single-molecule recognition through equilibrium Poisson sampling) analysis using one or more grayscale TIF stacks (movies) as input. The first program generates intensity-versus-time traces form grayscale TIF stacks for subsequent kinetic analysis. The second program performs kinetic analysis of the intensity-versus-time traces to determine whether each trace corresponds to nonspecific binding or specific binding to the target analyte, using a Parameters file to establish thresholds for this classification of traces. This method provides:
· Extremely low-background detection of single biomolecules (when probe design and analysis parameters are sufficiently optimized)
· Quantification with a dynamic range of typically ~2-3 orders of magnitude
· Ultra-high specificity capable of detecting single-base mutations in a 1 million-fold excess of wild-type sequence
See User Guide for more information, including links to example input and output files for testing.