IMPROVED INVASION ASSAY

摘要

The invention relates to improved methods of detecting and characterizing polynucleotide sequences and variations in polynucleotide sequences. The invention further relates to a device that can be used to detect and characterize polynucleotide sequences and variations in polynucleotide sequences. The invention is directed to an improved invasion assay to methods of analyzing invasion assay data. The invention is based, in part, on a discovery that signals due to the cleavage of a probe annealed to a template polynucleotide (e.g. DNA or RNA) can exhibit behavior over time that is different from the behavior over time of signals produced by background reactions (e.g. reactions that are not dependent on the presence of target polynuclotide). For example, the positive signal produced by many invasion assays (i.e., the signal that indicates an cleavage of a probe oligonucleotide annealed to a target polynucleotide) can be fit to a polynomial that is higher in order than the polynimial required to characterize the background signal. It is thus possible to determine the results of an invasion assay in real time or at the conclusion of the assay by determining whether the behavior of the signal it produced over time is different from that a typical, estimated theoretical or actually measured background signal.