This specification generally relates to non-radioactive methods of detecting nucleic acid polymerase activity and methods of detecting compounds that modulate nucleic acid polymerase activity. The activity may be measured in real-time using a real-time PCR instrument.

Disclosed are methods for isothermal nucleic acid amplification and detection.

Disclosed are methods for isothermal nucleic acid amplification and detection.

The invention provides comprehensive and comparative flow cytometry-based methods for characterizing the state of a biological system by determining cell phenotypes and associated gene expression profiles.

The invention provides comprehensive and comparative flow cytometry-based methods for characterizing the state of a biological system by determining cell phenotypes and associated gene expression profiles.

The present application relates to detection of changes in the number of nucleotides in short homopolymeric nucleic acid repeats, in particular in short homopolymeric microsatellites, for example for the purpose of diagnosing microsatellite instability (MSI) and/or mismatch repair (MMR-) deficiency in tumors. Accordingly, methods are provided for detecting changes in the number of nucleotides present in short homopolymeric nucleotide repeat sequences as well as kits and cartridges for automated detection of said changes.

The present application relates to detection of changes in the number of nucleotides in short homopolymeric nucleic acid repeats, in particular in short homopolymeric microsatellites, for example for the purpose of diagnosing microsatellite instability (MSI) and/or mismatch repair (MMR-) deficiency in tumors. Accordingly, methods are provided for detecting changes in the number of nucleotides present in short homopolymeric nucleotide repeat sequences as well as kits and cartridges for automated detection of said changes.

Compositions and methods for highly specific nucleic acid probes and primers are provided. The probe system comprises a complement strand and a protector stand that form a partially double-stranded probe. The reaction standard free energy of hybridization between the probe and target nucleic acid as determined by Expression 1 (ΔG°.sub.rxn=ΔG°.sub.t-TC−ΔG°.sub.nh-PC+(ΔG°.sub.v-TC−ΔG°.sub.h-PC)) is from about −4 kcal/mol to about +4 kcal/mol. Alternatively, the reaction standard free energy of hybridization between the probe and target nucleic acid is determined by Expression 1 to be within 5 kcal/mol of the standard free energy as determined by Expression 2 (−Rτln(([P].sub.0−[C].sub.0)/[C].sub.0)]), where the [P].sub.0 term of Expression 2 equals the concentration of the protector strand and the [C].sub.0 term of Expression 2 equals the concentration of the complement strand. In addition, a method for on-the-fly fine tuning of a reaction using the present probe is provided.

Compositions and methods for highly specific nucleic acid probes and primers are provided. The probe system comprises a complement strand and a protector stand that form a partially double-stranded probe. The reaction standard free energy of hybridization between the probe and target nucleic acid as determined by Expression 1 (ΔG°.sub.rxn=ΔG°.sub.t-TC−ΔG°.sub.nh-PC+(ΔG°.sub.v-TC−ΔG°.sub.h-PC)) is from about −4 kcal/mol to about +4 kcal/mol. Alternatively, the reaction standard free energy of hybridization between the probe and target nucleic acid is determined by Expression 1 to be within 5 kcal/mol of the standard free energy as determined by Expression 2 (−Rτln(([P].sub.0−[C].sub.0)/[C].sub.0)]), where the [P].sub.0 term of Expression 2 equals the concentration of the protector strand and the [C].sub.0 term of Expression 2 equals the concentration of the complement strand. In addition, a method for on-the-fly fine tuning of a reaction using the present probe is provided.

The present invention relates to methods and kits for determining the methylation status of a target nucleic acid molecule in a sample.