Patent ID: 6514700
Filing Date: 2003-02-04
Classification: C07H,C40B

Abstract:
A method for detecting at least one target nucleic acid sequence in a nucleic acid sample, said method comprising:combining said nucleic acid sample and at least one reagent pair consisting of a primer and an e-tag linked by a cleavable bond to a target-binding sequence, where at least one nucleotide linkage at positions immediately 3â€² of the second nucleotide at the 5â€²-end of said target-binding sequence is resistant to nuclease hydrolysis, each reagent pair having a sequence homologous to each nucleic acid sequence to be determined, wherein each said primer specifically binds to said target nucleic acid and said target-binding sequence binds to said target nucleic acid downstream from said primer, wherein each said target-binding sequence is characterized by being linked to a non-oligomeric e-tag specific for each said nucleic acid sequence; executing at least one cycle of cleavage of said cleavable bond, whereby said e-tag is released substantially free of said target-binding sequence, such that cleavage of the target-binding sequence at its 5â€² end produces a single released product comprised of the e tag and the 5â€²-end nucleotide of the target-binding sequence, wherein each said released product produced from a given target-binding sequence has a known, unique electrophoretic mobility with respect to the released products produced from all other such target-binding sequences, by virtue of a unique charge/mass ratio associated with the e tag; separating released products into individual fractions of released products specific for each target nucleic acid sequence; and detecting said released product fractions, whereby the presence in said target nucleic acid sample of said at least one nucleic acid sequence is detected.