Patent ID: 7081339
Filing Date: 2006-07-25
Classification: C12Q,G01N

Abstract:
1. A method for monitoring the amplification of a nucleic acid sequence of interest, said method comprising: (a) contacting a nucleic acid sample with a first and a second oligonucleotide primer, wherein said first oligonucleotide primer specifically hybridizes with a nucleic acid molecule comprising said nucleic acid sequence of interest, and said second oligonucleotide primer specifically hybridizes with a sequence comprised by the complement of said nucleic acid sequence of interest, wherein the primer extension product of one oligonucleotide primer, when separated from its complement, can serve as a template for the synthesis of the extension product of the other primer; (b) subjecting the mixture resulting from step (a) to an amplification regimen, said regimen comprising at least two cycles of nucleic acid strand separation, oligonucleotide primer annealing, and polymerase extension of annealed primers; (c) removing an aliquot of said mixture, separating nucleic acid molecules in said aliquot, and detecting amplified product, wherein said removing is performed during said cycling regimen of step (b), following at least one of said cycles, wherein said separating nucleic acid molecules and said detecting incorporation is performed in real time during said regimen of step (b) and wherein said detection permits the monitoring of said amplification in real time.