The present invention relates to a composition for DNA sequence analysis and a method for DNA sequence analysis, the method comprising treating a sample with the composition. The composition of the present invention can attain efficient optical identification at a single-DNA molecule level by linking both an A/T-specific DNA-binder agent and an A/T-non-specific complementary DNA-binder agent to DNA, and thus can be helpfully used in studying chromosomal organization of genomes, protein immunolocalization, and the like.

The present invention relates to a composition for DNA sequence analysis and a method for DNA sequence analysis, the method comprising treating a sample with the composition. The composition of the present invention can attain efficient optical identification at a single-DNA molecule level by linking both an A/T-specific DNA-binder agent and an A/T-non-specific complementary DNA-binder agent to DNA, and thus can be helpfully used in studying chromosomal organization of genomes, protein immunolocalization, and the like.

Methods are provided for nucleic acid analysis wherein a target nucleic acid that is at least partially double stranded is mixed with a dsDNA binding dye having a percent saturation of at least 50% to form a mixture. In one embodiment, the nucleic acid is amplified in the presence of the dsDNA binding dye, and in another embodiment a melting curve is generated for the target nucleic acid by measuring fluorescence from the dsDNA binding dye as the mixture is heated. Dyes for use in nucleic acid analysis and methods for making dyes are also provided.

Methods are provided for nucleic acid analysis wherein a target nucleic acid that is at least partially double stranded is mixed with a dsDNA binding dye having a percent saturation of at least 50% to form a mixture. In one embodiment, the nucleic acid is amplified in the presence of the dsDNA binding dye, and in another embodiment a melting curve is generated for the target nucleic acid by measuring fluorescence from the dsDNA binding dye as the mixture is heated. Dyes for use in nucleic acid analysis and methods for making dyes are also provided.

The present invention relates to the detection of a target nucleic acid sequence in a real-time manner using a target signal generating primer (TSG primer) having dual interactive labels. The present invention allows for both target amplification and signal amplification by introducing dual interactive labels into a primer used in PCR reactions, ensuring real-time target detection by PCR reactions without the use of complicated oligonucleotides. The present invention could be free from the troublesome matters and shortcomings associated with conventional real-time PCR methods. The present invention allows for successful real-time target detection by using only a labeled primer. Also, the present invention can obtain strong signals indicative of the presence of target nucleic acid sequences in both a liquid phase and solid phase.

The present invention relates to the detection of a target nucleic acid sequence in a real-time manner using a target signal generating primer (TSG primer) having dual interactive labels. The present invention allows for both target amplification and signal amplification by introducing dual interactive labels into a primer used in PCR reactions, ensuring real-time target detection by PCR reactions without the use of complicated oligonucleotides. The present invention could be free from the troublesome matters and shortcomings associated with conventional real-time PCR methods. The present invention allows for successful real-time target detection by using only a labeled primer. Also, the present invention can obtain strong signals indicative of the presence of target nucleic acid sequences in both a liquid phase and solid phase.

There is disclosed a kit for detecting a single strand target nucleotide sequence comprising: at least one first nucleic acid probe from 10 to 14 bases, to the 5′ end of which at least one fluorophore is bound; at least one second nucleic acid probe from 35 to 50 bases, comprising, from the 5′ to the 3′ end: a first segment having a nucleotide sequence complementary to the first nucleic acid probe, at least one quencher, and a second segment having a nucleotide sequence complementary to at least part of the target nucleotide sequence, wherein the following relation is met:
|ΔG hybr.target3−probe2|>|ΔG hybr.probe1−probe2|.

There is disclosed a kit for detecting a single strand target nucleotide sequence comprising: at least one first nucleic acid probe from 10 to 14 bases, to the 5′ end of which at least one fluorophore is bound; at least one second nucleic acid probe from 35 to 50 bases, comprising, from the 5′ to the 3′ end: a first segment having a nucleotide sequence complementary to the first nucleic acid probe, at least one quencher, and a second segment having a nucleotide sequence complementary to at least part of the target nucleotide sequence, wherein the following relation is met:
|ΔG hybr.target3−probe2|>|ΔG hybr.probe1−probe2|.

Provided herein are reagents and kits for detection of multiple target sequences in a single-tube, single-color assay, and methods of use thereof. In particular, multiplex assays are provided for the detection of Mycobacterium tuberculosis complex target sequences (e.g., katG, rpoB, inhA promotor, pncA, etc.).

Provided herein are reagents and kits for detection of multiple target sequences in a single-tube, single-color assay, and methods of use thereof. In particular, multiplex assays are provided for the detection of Mycobacterium tuberculosis complex target sequences (e.g., katG, rpoB, inhA promotor, pncA, etc.).