Abstract: The invention provides methods for the detection of the amount of a nucleic acid in a sample. The described methods exploit the ability to disrupt and redirect a PCR direction, and the ability to physically pair nucleic acid molecules in a sample that have a reference sequence with nucleic acid molecules in the sample that have a target sequence. The redirection of the PCR reaction enables partial amplification as a preparatory step to other techniques within the same tube. The pairing can result in the presence of unpaired target or reference sequence indicating a difference in the amount of the target sequence versus the reference sequence. The methods are broadly applicable for the determination of differences in the amount of nucleic acids in diagnostic and research applications.

Abstract: A method for detection of a quantitative difference between the amount of a first target region of nucleic acid and a second target region of nucleic acid in a sample, comprising the steps of: providing the sample comprising the nucleic acid; amplifying the first and second target regions of the nucleic acid to obtain multiple copies of a first and a second sequence of nucleic acid; associating the amplified first sequence with the amplified second sequence to form associated nucleic acid complexes which comprise the first sequence and the second sequence in a 1:1 ratio, wherein any excess of either the first sequence or the second sequence remain un-associated; detecting any un-associated sequences, wherein detection of any un-associated sequences is indicative of a quantitative difference between the amount of the first and second target regions of nucleic acid in the sample.