The present invention relates to a method and kit for amplifying and detecting a quantity of nucleic acid. The invention is particularly relevant to isothermal amplification techniques carried out on a flow based assay device. The amplified nucleic acid may be detected on the device using an optical read-out.

Provided are nucleic acid-based sensors, kits that include such sensors, and methods for making and using such sensors. The sensors permit detection of a broad array of target agents, such as nucleic acids (e.g., DNA and RNA), proteins, cells, and small molecules (e.g., toxins and metals). In some examples, the sensors determine the activity of a protein, such as an enzyme's activity. The assays provide easy-to-read signals for point-of-care diagnosis. The sensors control the conformational switch for recognizing targets and to trigger the subsequent assembly of enzyme/cofactor structures for producing and amplifying signals.

Provided are nucleic acid-based sensors, kits that include such sensors, and methods for making and using such sensors. The sensors permit detection of a broad array of target agents, such as nucleic acids (e.g., DNA and RNA), proteins, cells, and small molecules (e.g., toxins and metals). In some examples, the sensors determine the activity of a protein, such as an enzyme's activity. The assays provide easy-to-read signals for point-of-care diagnosis. The sensors control the conformational switch for recognizing targets and to trigger the subsequent assembly of enzyme/cofactor structures for producing and amplifying signals.

This disclosure related to methods of detecting mechanical forces required to separate ligand and receptor interactions. In certain embodiments, this disclosure relates to methods of detecting mechanical forces between a ligand and receptor, where the ligand is immobilized on a surface using weaker forces. Ligand-receptor forces lead to dissociation of the ligand that can be detected and amplified. In certain embodiments, the disclosure relates to methods of detecting ligand and receptor interactions comprising linking a ligand to one of two binding partners, wherein the binding partners have attracting forces that are less than the forces between the ligand and a receptor of the ligand such that when the ligand binds the receptor, the binding partners will separate. Separation of the binding partners can be detected.

This disclosure related to methods of detecting mechanical forces required to separate ligand and receptor interactions. In certain embodiments, this disclosure relates to methods of detecting mechanical forces between a ligand and receptor, where the ligand is immobilized on a surface using weaker forces. Ligand-receptor forces lead to dissociation of the ligand that can be detected and amplified. In certain embodiments, the disclosure relates to methods of detecting ligand and receptor interactions comprising linking a ligand to one of two binding partners, wherein the binding partners have attracting forces that are less than the forces between the ligand and a receptor of the ligand such that when the ligand binds the receptor, the binding partners will separate. Separation of the binding partners can be detected.

Kits, compositions, and methods for labeling target materials or target analytes are considered. The composition can include one or more affinity molecules indirectly conjugated to one or more detection moieties. The kit can be used to detect a single biomarker or multiple biomarkers. The kit can be used to perform a single round of labeling or multiple rounds of labeling.

Kits, compositions, and methods for labeling target materials or target analytes are considered. The composition can include one or more affinity molecules indirectly conjugated to one or more detection moieties. The kit can be used to detect a single biomarker or multiple biomarkers. The kit can be used to perform a single round of labeling or multiple rounds of labeling.

Compositions and methods comprising the use of polymerase-nucleotide conjugates for nucleic acid sequencing and analysis applications are described.

Compositions and methods comprising the use of polymerase-nucleotide conjugates for nucleic acid sequencing and analysis applications are described.

This disclosure provides a method of determining a sequence of nucleotides for a nucleic acid template. The method can include the steps of contacting the nucleic acid template with a conformationally labeled polymerase and at least four different nucleotide species under conditions wherein the conformationally labeled polymerase catalyzes sequential addition of the nucleotide species to form a nucleic acid complement of the nucleic acid template, wherein the sequential addition of each different nucleotide species produces a conformational signal change from the conformationally labeled polymerase and wherein the rate or time duration for the conformational signal change is distinguishable for each different nucleotide species; detecting a series of changes in the signal from the conformationally labeled polymerase under the conditions; and determining the rates or time durations for the changes in the signal, thereby determining the sequence of nucleotides for the nucleic acid template.