Patent ID: 8288093

Claim:
A method comprising: a) providing: i) a first complex comprising a first nucleic acid, a second nucleic acid and a third nucleic acid, said first nucleic acid comprising a first portion and said second nucleic acid comprising a first region and a second region, wherein said first region of said second nucleic acid is 5â€² of said second region of said second nucleic acid, and wherein said first portion of said first nucleic acid is hybridized to said first region of said second nucleic acid, and wherein at least a portion of said third nucleic acid is hybridized to said second region of said second nucleic acid; and ii) a fourth nucleic acid comprising a hairpin structure and a detectable label; b) cleaving said first complex thereby generating a first cleavage product, wherein the amount of said first cleavage product increases at a first rate, and increases as a function of time; c) hybridizing at least a portion of said first cleavage product to at least a portion of said fourth nucleic acid thereby forming a second complex; d) cleaving said fourth nucleic acid in said second complex thereby generating a second cleavage product comprising said detectable label, wherein the amount of said second cleavage product increases at a second rate, and increases as a function of time; and e) detecting the amount of said second cleavage product; and f) determining said second rate.