Abstract: A method for determining the presence, absence or amount of two or more target polynucleotides in a sample comprising additional components, the method comprising: (i) contacting the sample with a panel of two or more probes under conditions suitable for hybridisation of the target polynucleotides to the probes, wherein: (a) each probe comprises a non-hybridisation region and a hybridisation region that specifically hybridises to one of the target polynucleotides to form a hybridised probe; and (b) the hybridisation region of a probe of the panel comprises one or more non-natural nucleotides; (ii) contacting the sample prepared in step (i) with a transmembrane pore through which a single stranded polynucleotide but not a double stranded polynucleotide can pass and applying a potential difference to the transmembrane pore such that the hybridised probes in the sample interact with the pore; (iii) measuring current blockades having a duration within a defined window, wherein: (a) the one or more non-natural nuc

Abstract: The present disclosure relates to base sequence determination. A base sequence determination system includes a fluid device and an optical device, a reaction device includes a first component and a second component, and a repeated executable unit is defined as: a second biochemical reaction—a first biochemical reaction—photographing. A method includes, after initiation steps, using the fluid device to perform the second biochemical reaction and the first biochemical reaction of the sample on the first component, while using the optical device to photograph the sample on the second component. The initial steps include: using the fluid device to perform the first biochemical reaction of the sample on the first component, using the optical device to photograph the sample on the first component after the first biochemical reaction, and using the fluid device to perform the first biochemical reaction of the sample on the second component.

Abstract: The present invention provides assays and assay systems for use in spatially encoded biological assays. The invention provides an assay system comprising an assay capable of high levels of multiplexing where reagents are provided to a biological sample in defined spatial patterns; instrumentation capable of controlled delivery of reagents according to the spatial patterns; and a decoding scheme providing a readout that is digital in nature.

Abstract: The nucleotides can then be polymerized into oligomers. The design of the oligomers will depend on the design of the overall architecture. Simple architectures may be designed by any methods. However, more complex architectures may be design using software, such as caDNAno (as described at cadnano.org/docs.html, and herein incorporated by reference), to minimize errors and time. The user may input the desired shape of the architecture into the software and once finalized, the software will provide the oligomer sequences of the bricks to create the desired architecture. The length of the oligomers may be from about 10 to about 10,000, or less than about 9,000, less than about 8,000, less than about 5,000 nucleotides in length. The length of the oligomer will be optimized for the type of architecture used.

Abstract: The present invention provides assays and assay systems for use in spatially encoded biological assays. The invention provides an assay system comprising an assay capable of high levels of multiplexing where reagents are provided to a biological sample in defined spatial patterns; instrumentation capable of controlled delivery of reagents according to the spatial patterns; and a decoding scheme providing a readout that is digital in nature.

Abstract: The present application relates to detection units and methods for detecting one or more target analytes in a sample using a complex formed by a target and first and second probes, wherein the first probe is coupled to a detectable piece, the target is coupled to the first probe and the second probe, and the second probe is coupled to a solid support. Specific binding of the detectable piece to the target analyte can be distinguished from non-specific binding of the detectable piece by measuring the number of detectable pieces that leave their initial location after exposure to a disruptor that uncouples the detectable piece from the solid support.

Abstract: In the present invention, a fraction (A) is labeled, the fraction (A) being a fraction obtained from a maternal blood sample and in which nucleated red blood cells (NRBCs) are concentrated in a population of whole blood cells. Then, a fraction (B) having increase purity of NRBCs is obtained by sorting out blood cells in the labeled fraction A by at least cell sorting. Next, fractions (C) are obtained by separating each blood cell in the fraction (B) at a single-cell level and independently performing a process for extracting a nucleic acid for each separated blood cell, each of the fractions (C) containing a nucleic acid distinguishable at a single-cell level. Then, a fraction (D) containing a nucleic acid derived from a fetus is sorted out from a group of fractions (C) by performing a molecular biological analysis for each of the fractions (C).

Abstract: The present invention relates to methods and systems that result in high quality, reproducible, thermal melt analysis on a microfluidic platform. The present invention relates to methods and systems using thermal systems including heat spreading devices, including interconnection methods and materials developed to connect heat spreaders to microfluidic devices. The present invention also relates to methods and systems for controlling, measuring, and calibrating the thermal systems of the present invention.

Abstract: The present disclosure provides methods and systems for nucleic acid preparation and analysis. Nucleic acid molecules may be derived from one or more cells. Preparation of nucleic acid molecules may comprise generation of one or more mutations, such as strand-specific mutations. Nucleic acid molecules may be prepared for and analyzed by sequencing. Sequences may be identified with nucleic acid orientation information.

Abstract: The present disclosure provides methods and assay systems for use in spatially encoded biological assays, including assays to determine a spatial pattern of abundance, expression, and/or activity of one or more biological targets across multiple sites in a sample. In particular, the biological targets comprise proteins, and the methods and assay systems do not depend on imaging techniques for the spatial information of the targets. The present disclosure provides methods and assay systems capable of high levels of multiplexing where reagents are provided to a biological sample in order to address tag the sites to which reagents are delivered; instrumentation capable of controlled delivery of reagents; and a decoding scheme providing a readout that is digital in nature.

Abstract: Methods are provided for detecting and quantitating molecules using fluidics. In preferred embodiments, the methods comprise analyzing blood to detect the presence of circulating DNA or cells from a fetus or tumor.

Abstract: Disclosed herein are methods of detecting a target nucleic acid sequence, determining the localization of the target nucleic acid sequence, and/or quantifying the number of target nucleic acid sequences in a cell. This method may be used on small target nucleic acid sequences, and may be referred to as Nano-FISH.

Abstract: Disclosed is a structure for detecting cell-free DNA using a conductive polymer and the use thereof. More particularly, a nanostructure composed of a surface-modified conductive polymer for isolating and detecting cell-free DNA is disclosed. By using the nanostructure, cell-free DNA including circulating tumor DNA, can be effectively detected and isolated.

Abstract: The invention provides methods for detecting specific nucleic acids using a loop-mediated isothermal amplification (LAMP) reaction and providing a digital readout of the results. One method teaches separating target specificity from indicator detection in a two-stage multiple-threshold array (MTA). During the first stage amplicons incorporating both target sequence and an indicator sequence are synthesized. During the second stage of the reaction, the indicator sequence is targeted and amplified to produce visual results that may be digitalized. Another method teaches a competitive MTA, in which a reference sequence is used to compete against target sequence during amplification producing threshold responses, thus enabling digital readout. Using either method, quantitation is achieved without the need for continuous monitoring of the reaction and the end-point readout is amenable to visual inspection.

Abstract: Provided herein are methods, systems, and compositions for seamless nucleic acid assembly. Methods, systems, and compositions as provided herein provide for efficient assembly of nucleic acids without primer removal. Methods, systems, and compositions for seamless nucleic acid assembly comprise use of an endonuclease or exonuclease, optionally in conjunction with additional enzymes to assemble nucleic acids or polynucleotides.

Abstract: Disclosed herein are detection devices, systems including such detection devices, and methods of using such detection devices. A detection device comprises a fluidic channel configured to receive a plurality of molecules to be detected, a plurality of temperature sensors, and an insulating material encapsulating the plurality of temperature sensors and providing a barrier between the plurality of temperature sensors and contents of the fluidic channel. A surface of the insulating material within the fluidic channel provides a plurality of sites for binding the plurality of molecules to be detected. Each of the plurality of temperature sensors is configured to detect, in the presence of an alternating magnetic field, a temperature change indicating presence or absence of one or more magnetic nanoparticles (MNPs) coupled to at least one of the plurality of molecules to be detected at a respective subset of the plurality of sites.

Abstract: The present disclosure provides methods and systems for nucleic acid sequencing. Such systems and methods may achieve context-independent incorporation, have reduced context-dependence or have context-dependence that is amenable to calibration and modeling. Such systems and methods may also reduce misincorporation.

Abstract: Provided herein are methods for determining presence of a target enzyme in a tissue section that include delivering a plurality of probes to a tissue section, where a probe of the plurality of probes comprises a capture agent that comprises a substrate for the target enzyme in the tissue section, and where the capture agent is conjugated to an oligonucleotide.

Abstract: Disclosed herein are compositions for fixing tissue for cytologic, histomorphologic, and/or molecular analysis (e.g., DNA, RNA, and/or protein analysis). In some embodiments, the fixatives are aldehyde-free fixatives, for example, formaldehyde- or formalin-free fixatives. Particular disclosed compositions include buffered ethanol. The buffer is a phosphate buffer or phosphate buffered saline (PBS) in some examples. In further embodiments, the fixative includes additional components, such as glycerol and/or acetic acid.

Abstract: Systems and methods are provided herein for encoding and storing information in nucleic acids. Encoded information is partitioned and stored in nucleic acids having native key-value pairs that allow for storage of metadata or other data objects. Computation on the encoded information is performed by chemical implementation of if-then-else operations. Numerical data is stored in nucleic acids by producing samples having nucleic acid sequences copy counts corresponding to the numerical data. Data objects of a dataset are encoded by partitioning of bytes into parts and encoding of parts along distinct libraries of nucleic acids. These libraries can be used as inputs for computation on the dataset.