Patent ID: 6306607
Filing Date: 2001-10-23
Classification: C07D,C07H,C12Q,Y10T

Abstract:
A heterogeneous assay method for detecting pyrophosphate cleavage, the components of the assay comprising a labeled NTP, a target nucleic acid, a primer nucleic acid and a polymerase, said method comprising:(a) flowing said labeled nucleotide triphosphate (NTP) having a .gamma.-phosphate with a fluorophore moiety attached thereto, past an immobilized component selected from the group consisting of said polymerase and said target nucleic acid;(b) incorporating said NTP on a primer strand hybridized to said target nucleic acid using said polymerase and releasing said .gamma.-phosphate with said fluorophore moiety attached thereto; and(c) detecting said fluorescent moiety thereby detecting pyrophosphate cleavage.