A cleavage-based real-time PCR assay method is provided. In general terms, the assay method includes subjecting a reaction mixture comprising a) PCR reagents for amplifying a nucleic acid target, and b) flap cleavage reagents for performing a flap cleavage assay on the amplified nucleic acid target to two sets of thermocycling conditions. No additional reagents are added to the reaction between said first and second sets of cycles and, in each cycle of the second set of cycles, cleavage of a flap probe is measured.

A cleavage-based real-time PCR assay method is provided. In general terms, the assay method includes subjecting a reaction mixture comprising a) PCR reagents for amplifying a nucleic acid target, and b) flap cleavage reagents for performing a flap cleavage assay on the amplified nucleic acid target to two sets of thermocycling conditions. No additional reagents are added to the reaction between said first and second sets of cycles and, in each cycle of the second set of cycles, cleavage of a flap probe is measured.

Provided herein is technology relating to the amplification-based detection of bisulfite-treated DNAs and particularly, but not exclusively, to methods and compositions for multiplex amplification of low-level sample DNA prior to further characterization of the sample DNA. The technology further provides methods for isolating DNA from blood or blood product samples, e.g., plasma samples.

Provided herein is technology relating to the amplification-based detection of bisulfite-treated DNAs and particularly, but not exclusively, to methods and compositions for multiplex amplification of low-level sample DNA prior to further characterization of the sample DNA. The technology further provides methods for isolating DNA from blood or blood product samples, e.g., plasma samples.

A probe system for real-time quantitative and qualitative analysis of a biomaterial, and a reaction chamber with the probe system, and an analysis method thereof are provided. The probe system, which is included in the reaction chamber having an optically transmissive flat bottom surface and having a test sample accommodated therein, includes a target probe-reporter probe linker accommodated in the reaction chamber and including a target probe, which includes a sequence complementary to a target nucleic acid sequence to be detected, a first fluorophore and a first quencher, and a reporter probe linked to an end of the target probe and including a sequence non-complementary to the target nucleic acid sequence, and a capture probe included in a biochip formed on a bottom surface of the reaction chamber and including a complementary sequence hybridizable with the non-complementary sequence of the reporter probe, a second fluorophore and a second quencher.

A probe system for real-time quantitative and qualitative analysis of a biomaterial, and a reaction chamber with the probe system, and an analysis method thereof are provided. The probe system, which is included in the reaction chamber having an optically transmissive flat bottom surface and having a test sample accommodated therein, includes a target probe-reporter probe linker accommodated in the reaction chamber and including a target probe, which includes a sequence complementary to a target nucleic acid sequence to be detected, a first fluorophore and a first quencher, and a reporter probe linked to an end of the target probe and including a sequence non-complementary to the target nucleic acid sequence, and a capture probe included in a biochip formed on a bottom surface of the reaction chamber and including a complementary sequence hybridizable with the non-complementary sequence of the reporter probe, a second fluorophore and a second quencher.

Methods and compositions for the detection and quantification of nucleic acids are provided. In certain embodiments, methods involve the use of primers or probes that comprise a non-natural nucleotide linked to a reporter. Target nucleic acids are detected by the polymerization of a complementary probe or primer that incorporated a cognate non-natural nucleotide linked to a quencher.

Methods and compositions for the detection and quantification of nucleic acids are provided. In certain embodiments, methods involve the use of primers or probes that comprise a non-natural nucleotide linked to a reporter. Target nucleic acids are detected by the polymerization of a complementary probe or primer that incorporated a cognate non-natural nucleotide linked to a quencher.

Provided herein is reagent mixture comprising multiplexed amplification reagents and flap assay reagents for detecting, in a single reaction, mutant copies of the KRAS gene that contain any of the 34A, 34C, 34T, 35A, 35C, 35T or 38A point mutations. Methods that employ the reagent mix and kits for performing the same are also provided.

Provided herein is reagent mixture comprising multiplexed amplification reagents and flap assay reagents for detecting, in a single reaction, mutant copies of the KRAS gene that contain any of the 34A, 34C, 34T, 35A, 35C, 35T or 38A point mutations. Methods that employ the reagent mix and kits for performing the same are also provided.