monitoring known mutations invasive cleavage invader SISAR MIRO:20000123 MIRO:20000123 The method requires annealing of two oligonucleotides, called the upstream oligonucleotide and the probe, to a target sequence, which results in the formation of a unique substrate for the 5' nuclease. The probe contains two regions, an analyte-specific region that forms a duplex with the target and a noncomplementary 5' arm region, which is not required for enzyme activity but serves as a reporter molecule precursor. Cleavage of the probe occurs only when the probe and upstream oligonucleotide overlap; therefore, two target DNAs differing only by a single nucleotide that affects formation of the cleavage structure can be differentiated. This extraordinary specificity of substrate structure recognition by the 5' nuclease enables detection of single point mutations with a discrimination level required for single-nucleotide polymorphism analysis. mosquito_insecticide_resistance_ontology