Abstract: A novel method for characterizing nucleic acids. A nucleic acid is combined with a double stranded nucleic acid-specific dye to form a detectable complex between the dye and one or more double stranded structures within the nucleic acid. The combination is then exposed to varying temperatures and the fluorescence emission of the dye is measured to determine the melting temperature(s) for the double stranded structures. In some embodiments that melting temperature profile is then compared to melting temperature profiles generated for other nucleic acid(s) to discern differences between the compared nucleic acids.

Abstract: The present invention is directed to kits for monitoring a nucleic acid during amplification. More particularly, the present invention relates to kits comprising a first oligonucleotide labeled with a first fluorophore, a second oligonucleotide labeled with a second fluorophore, and components for amplifying the locus, wherein the first and second fluorophores comprise a fluorescence resonance energy transfer pair. Hybridization of at least the first oligonucleotide to the amplified locus places the first and second fluorophores in a resonance energy transfer relationship.

Abstract: Methods for analyzing a target nucleic acid are provided. A fluorescent label attached to a nucleic acid is incorporated into at least one strand of the target nucleic acid and the methods include monitoring change in fluorescence emission resulting from dissociation of the labeled strand of the amplification product from its complementary strand.

Abstract: Probes and methods are provided for detection and analysis of nucleic acid sequences. The probes are single-labeled oligonucleotide probes whose fluorescence emission changes in response to probe-target hybridization and dissociation. The methods are for analyzing one or multiple nucleic acid loci using the probes. This invention further relates to the use of fluorescence changes in single-labeled probes for melting curve analysis, genotyping, and pathogen detection, and to methods for quantification of specific sequences in real-time monitoring of nucleic acid amplification.

Abstract: A method and device are described for analyzing a sample for the presence of a nucleic acid wherein the sample is amplified, illustratively using PCR, in the presence of a fluorescent probe capable of providing a signal related to the quantity of nucleic acid present. A nucleic acid sample is amplified in the presence of the fluorescent entity, and the fluorescence intensity is measured at each of a plurality of amplification cycles. Scores are obtained from various tests performed on the fluorescence data, and the scores are used to determine whether the nucleic acid is present in the sample.

Abstract: A method and device are described for analyzing a sample for the presence of an analyte wherein the analyte is contacted with a substrate to effect a measurable change selected from the group consisting of the quantity of the analyte, the quantity of the substrate, and the quantity of an optical or physical change to the substrate, wherein the analyte is contacted with the substrate for a predetermined time period, to generate a signal related to the measurable change. Scores are obtained from various tests performed on the signal data, and the scores are used to determine whether the substrate is present in the sample.

Abstract: The present invention is directed to a method and kits for monitoring a nucleic acid during amplification. More particularly, the present invention relates to a method wherein the nucleic acid is monitored during polymerase chain reaction using a double-stranded nucleic acid binding dye capable of producing a fluorescent signal related to the amount of the nucleic acid present in a sample, wherien the dye is selected from the group consisting of SYBR™ Green I and pico green.

Abstract: Probes and methods are provided for detection and analysis of nucleic acid sequences. The probes are single-labeled oligonucleotide probes whose fluorescence emission changes in response to probe-target hybridization and dissociation. The methods are for analyzing one or multiple nucleic acid loci using the probes. This invention further relates to the use of fluorescence changes in single-labeled probes for melting curve analysis, genotyping, and pathogen detection, and to methods for quantification of specific sequences in real-time monitoring of nucleic acid amplification.

Abstract: Methods are provided for quantifying the concentration of a nucleic acid in a nucleic acid sample. The methods include contacting the nucleic acid sample with an amplifying agent, amplifying at least one predetermined locus of the nucleic acid by subjecting the sample to a number of amplification, generating an amplification curve or array, calculating the first, second or n th order derivative of the amplification curve or array, determining a maximum, minimum, or zero value of the derivative, and using the maximum, minimum, or zero value to calculate the initial concentration of the nucleic acid in the nucleic acid sample.

Abstract: The invention provides methods and devices for analyzing sequence variations in nucleic acid samples comprising multiple loci, each having two, three or more possible allelic sequences. The method involves combining at least a first and second pair of oligonucleotide probes with the nucleic acid sample. The first pair of probes is capable of hybridizing in proximity to each other within a segment of the nucleic acid sample comprising the first locus and the second pair is capable of hybridizing in proximity to each other within a segment of the nucleic acid sample comprising the second locus. The first member of each probe pair comprises a FRET donor and the second member comprises a FRET acceptor, the FRET acceptor of the first probe pair member having a different emission spectrum from the FRET acceptor of the second probe pair.

Abstract: A system for thermal cycling a sample utilizing electrolyte conductance heating. An electric current is passed through the sample to increase its temperature due to resistance heating. As the sample acquires more thermal energy its viscosity changes, which causes a change in resistance. Because of this characteristic, temperature of the sample can be measured as a function of resistance and temperature can be controlled using resistance of the solution as feedback to a circuit which controls the heating of the sample by electrical conductance.

Abstract: Methods for identifying and locating alterations in a nucleic acid having a known sequence are provided. The methods involve measuring the melting temperature of probe nucleic acids hybridized to a target nucleic acid. The methods take advantage of the differential dissociation temperatures of a probe from a target resulting from mismatches at different locations along the region of the target to which the probe hybridizes.

Abstract: A method and device are described for analyzing a sample for the presence of a nucleic acid wherein the sample is amplified, illustratively using PCR, in the presence of a fluorescent probe capable of detecting the presence of the nucleic acid sample. A baseline region is determined by comparing the fluorescence at various amplification cycles, and the fluorescence at a selected amplification cycle is compared to the baseline region to determine whether the fluorescence measurement falls outside of that baseline region, indicating the presence of the nucleic acid. A positive result may be verified by melting temperature analysis.

Abstract: The present invention is directed to a method and kits for monitoring a nucleic acid during amplification. More particularly, the present invention relates to a method wherein the nucleic acid is monitored during polymerase chain reaction using a heat-labile double-stranded nucleic acid binding dye capable of producing a fluorescent signal related to the amount of the nucleic acid present in a sample.

Abstract: A method is described for analyzing a sample for the presence of a nucleic acid wherein the sample is amplified, illustratively using PCR, in the presence of a fluorescent probe capable of detecting the presence of the nucleic acid sample. A baseline region is determined by comparing the fluorescence at various amplification cycles, and the fluorescence at a selected amplification cycle is compared to the baseline region to determine whether the fluorescence measurement falls outside of that baseline region, indicating the presence of the nucleic acid.

Abstract: Methods are provided for quantifying the concentration of a nucleic acid in a nucleic acid sample. The methods include contacting the nucleic acid sample with an amplifying agent, amplifying at least one predetermined locus of the nucleic acid by subjecting the sample to a number of amplification, generating an amplification curve or array, calculating the first, second or n th order derivative of the amplification curve or array, determining a maximum, minimum, or zero value of the derivative, and using the maximum, minimum, or zero value to calculate the initial concentration of the nucleic acid in the nucleic acid sample.

Abstract: The invention provides a new method for the quantification of a nucleic acid, wherein in a first step, the nucleic acid is amplified by an amplifying agent. Subsequently, first, second or n th order derivatives is calculated. The obtained value can then be used to calculate the initial concentration of the analyte.

Abstract: A thermal cycling method and device is disclosed. The device comprises a sample chamber whose temperature can be rapidly and accurately modulated over a range of temperatures needed to carry out a number of biological procedures, such a the DNA polymerase chain reaction. Biological samples are placed in glass micro capillary tubes and then located inside the sample chamber. A programmable controller regulates the temperature of the sample inside the sample chamber. Once a heating cycle is completed, the controller opens a door to the chamber for venting hot air out and cool ambient air is moved in. Temperature versus time profiles corresponding to optimum denaturation, annealing and elongation temperatures for amplification of DNA are achieved by the present invention.

Abstract: The present invention relates to fluorescence resonance energy transfer pairs for detecting the presence of a target analyte wherein the donor fluorophore's emission spectrum and the acceptor fluorophore's absorption spectrum overlap by less than 25%. In a preferred embodiment, the present invention relates to the use of fluorescein and Cy5 or Cy5.5 as fluorescence resonance energy transfer pairs for use as labels on oligonucleotides for analysis of a nucleic acid locus during amplification.

Abstract: The present invention is directed to a method of determining the concentration of a nucleic acid product that had been amplified through polymerase chain reaction (PCR). More particularly, the present invention relates to a method wherein a rate constant is determined for a known concentration of amplified product by monitoring the rate of hybridization of the known concentration, and then the concentration of an unknown concentration of a nucleic acid product can be determined by determining the rate of annealing for the unknown concentration, and calculating the concentration from the rate of annealing and the rate constant.