Described is an assay termed Programmable Enzyme-Assisted Selective Exponential Amplification (PASEA) that concurrently amplifies both wild type and mutant alleles while selectively cleaving the former. With time, the rare mutant alleles dominate, and are readily detectable by direct detection, Sanger sequencing, and other readily available methods. Also described are point-of-care assays and microfluidic devices for performing PASEA.

Described is an assay termed Programmable Enzyme-Assisted Selective Exponential Amplification (PASEA) that concurrently amplifies both wild type and mutant alleles while selectively cleaving the former. With time, the rare mutant alleles dominate, and are readily detectable by direct detection, Sanger sequencing, and other readily available methods. Also described are point-of-care assays and microfluidic devices for performing PASEA.

A method for analyzing planar sample is provided. In some cases the method comprises: (a) incubating the planar sample with a capture agent that is linked to an oligonucleotide, wherein the capture agent specifically binds to complementary sites in the planar sample; (b) reading a fluorescent signal caused by extension of a primer that is hybridized to the oligonucleotide, using fluorescence microscopy. Several implementations of the method, and multiplexed versions of the same, are also provided.

A method for analyzing planar sample is provided. In some cases the method comprises: (a) incubating the planar sample with a capture agent that is linked to an oligonucleotide, wherein the capture agent specifically binds to complementary sites in the planar sample; (b) reading a fluorescent signal caused by extension of a primer that is hybridized to the oligonucleotide, using fluorescence microscopy. Several implementations of the method, and multiplexed versions of the same, are also provided.

Provided herein are compositions and methods for sequencing using at least altering electrical characteristics of polymer ridges. In some examples, the bridges may span the space between first and second electrodes and may include a single-stranded conjugated polymer chain. A plurality of nucleotides may be coupled to corresponding labels. A polymerase may be coupled to the bridge and may add nucleotides to a first polynucleotide using at least a sequence of a second polynucleotide. The labels corresponding to those nucleotides respectively may alter an electrical characteristic of the conjugated polymer chain. Detection circuitry may detect a sequence in which the polymerase adds the nucleotides to the first polynucleotide using at least changes in an electrical signal through the bridge, the changes being responsive to the respective alterations of hybridization using the labels corresponding to those nucleotides.

Provided herein are compositions and methods for sequencing using at least altering electrical characteristics of polymer ridges. In some examples, the bridges may span the space between first and second electrodes and may include a single-stranded conjugated polymer chain. A plurality of nucleotides may be coupled to corresponding labels. A polymerase may be coupled to the bridge and may add nucleotides to a first polynucleotide using at least a sequence of a second polynucleotide. The labels corresponding to those nucleotides respectively may alter an electrical characteristic of the conjugated polymer chain. Detection circuitry may detect a sequence in which the polymerase adds the nucleotides to the first polynucleotide using at least changes in an electrical signal through the bridge, the changes being responsive to the respective alterations of hybridization using the labels corresponding to those nucleotides.

In an amplification reaction in a microfluidic apparatus, the reaction is carried out using starting substances tagged with fluorophore and quencher. The detection of reaction products occurs according to the disclosure by a separation of fluorophore and quencher occurring in the context of the amplification reaction. For the detection reaction, at least one energy-transferring substance is added and the evaluation occurs on the basis of the fluorescence emission of the fluorophores which occurs.

In an amplification reaction in a microfluidic apparatus, the reaction is carried out using starting substances tagged with fluorophore and quencher. The detection of reaction products occurs according to the disclosure by a separation of fluorophore and quencher occurring in the context of the amplification reaction. For the detection reaction, at least one energy-transferring substance is added and the evaluation occurs on the basis of the fluorescence emission of the fluorophores which occurs.

This disclosure provides for methods and reagents for rapid multiplex RPA reactions and improved methods for detection of multiplex RPA reaction products. In addition, the disclosure provides new methods for eliminating carryover contamination between RPA processes.

This disclosure provides for methods and reagents for rapid multiplex RPA reactions and improved methods for detection of multiplex RPA reaction products. In addition, the disclosure provides new methods for eliminating carryover contamination between RPA processes.