The disclosure provides methods and systems for nucleic acid amplification including isothermal nucleic acid amplification.

The disclosure provides methods and systems for nucleic acid amplification including isothermal nucleic acid amplification.

The present disclosure provides, in some aspects, nucleic acid-based molecular tools that enable the recording of molecular structure and soluble signals as well as the programmed assembly of molecular structures.

The present disclosure provides, in some aspects, nucleic acid-based molecular tools that enable the recording of molecular structure and soluble signals as well as the programmed assembly of molecular structures.

A nucleic acid sequence measuring method includes measuring fluorescence from the nucleic acid sequence measuring device supplied with a sample solution. The device includes a fluorescent probe added with a fluorescent molecule, and a quenching probe added with a quenching substance. The fluorescent probe and/or the quenching probe has a detection part detecting a predetermined nucleic acid sequence. Fluorescence from the fluorescent molecule is quenched by the quenching substance coupled with the fluorescent molecule when the hybridization between the detection target nucleic acid and the detection part has not occurred, and fluorescence is emitted from the fluorescent molecule separated from the quenching substance when the hybridization has occurred.

A nucleic acid sequence measuring method includes measuring fluorescence from the nucleic acid sequence measuring device supplied with a sample solution. The device includes a fluorescent probe added with a fluorescent molecule, and a quenching probe added with a quenching substance. The fluorescent probe and/or the quenching probe has a detection part detecting a predetermined nucleic acid sequence. Fluorescence from the fluorescent molecule is quenched by the quenching substance coupled with the fluorescent molecule when the hybridization between the detection target nucleic acid and the detection part has not occurred, and fluorescence is emitted from the fluorescent molecule separated from the quenching substance when the hybridization has occurred.

Probe systems and methods are provided for detecting nucleic acid targets using labeled polynucleotide probes and antiprobes that interact together and with complementary targets. These interactions result in signaling changes that indicate target frequency and provide error-checking functions that facilitate single base discrimination. These probe:antiprobe compositions enable real-time PCR detection, end-point detection and microarray detection of microbial species, drug resistant mutants, and cancer related variants. The probe:antiprobe may be an internal probe between two primers or may be a primer-probe. The probe also may be modified by introducing a base mismatch to increase thermodynamic discrimination of a correct versus incorrect target differing by a single base. Probe systems also are provided for use in methods of increasing target amplification and detecting specific single base variants.

Probe systems and methods are provided for detecting nucleic acid targets using labeled polynucleotide probes and antiprobes that interact together and with complementary targets. These interactions result in signaling changes that indicate target frequency and provide error-checking functions that facilitate single base discrimination. These probe:antiprobe compositions enable real-time PCR detection, end-point detection and microarray detection of microbial species, drug resistant mutants, and cancer related variants. The probe:antiprobe may be an internal probe between two primers or may be a primer-probe. The probe also may be modified by introducing a base mismatch to increase thermodynamic discrimination of a correct versus incorrect target differing by a single base. Probe systems also are provided for use in methods of increasing target amplification and detecting specific single base variants.

The subject invention pertains to composition and methods of using said composition as an in vitro biosensor of small molecules in biological and/or environmental samples using enzyme-assisted nucleic acid reactions. The methods and compositions can be used to sense and/or transduce the signal of a sensing event mediated by allosteric proteins, endonucleases and nucleic acid reactions. This invention allows the rapid development and setup of one-pot assays to provide results in minutes. The methods and compositions may be used to generate an electrochemical, fluorescent, colorimetric, and/or luminescent output and the methods can be performed in different modalities, including a solution-based or paper-based assay.

The subject invention pertains to composition and methods of using said composition as an in vitro biosensor of small molecules in biological and/or environmental samples using enzyme-assisted nucleic acid reactions. The methods and compositions can be used to sense and/or transduce the signal of a sensing event mediated by allosteric proteins, endonucleases and nucleic acid reactions. This invention allows the rapid development and setup of one-pot assays to provide results in minutes. The methods and compositions may be used to generate an electrochemical, fluorescent, colorimetric, and/or luminescent output and the methods can be performed in different modalities, including a solution-based or paper-based assay.