Assaying nucleotides in solution using a fluorescent intensity quenching effect
AssigneeLorne Park Research, Inc.
InventorsYuan Min Wu, Eileen Xiao-Feng Nie
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Abstract
The invention provides a method for rapidly, economically and efficiently sequencing and assaying nucleotides in a liquid medium using laser induced fluorescence of antisense probes, including PNA probes. Fluorescent intensity of the resulting medium is inversely proportional to the hybridization efficiency of the probes with respect to the target sequence. The method is particularly advantageous in not requiring separation of unhybridized probes and hybridization complexes prior to detection. The method can be used to identify accessible regions in folded nucleotide sequences, to determine the number of mismatched pairs in a hybridization complex, and to map genomes.
Claims28
A method for detecting at least one single stranded or double stranded nucleotide sequence in a liquid medium, said method comprising:
- providing said liquid medium comprising said at least one nucleotide sequence;
- adding to said liquid medium at least one antisense probe to form at least one hybridization complex with said at least one nucleotide sequence, wherein said at least one probe comprises at least one marker;
- and
- detecting said at least one nucleotide sequence by detecting at least one signal that is inversely proportional to an amount of said at least one hybridization complex in said liquid medium and proportional to an amount of said at least one probe unhybridized in said liquid medium,
- wherein said method is conducted without separating unhybridized probes from said at least one hybridization complex prior to said signal detecting, and without providing a signal quenching agent on said at least one probe or on said at least one nucleotide sequence.
Dependencies are inferred from the claim text.