Abstract: A method for analyzing macromolecules, including peptides, polypeptides, and proteins, employing nucleic acid encoding is disclosed.

Abstract: Disclosed is a method of fragmenting DNA comprising contacting a sample of target DNA with (a) a composition comprising an active transpososome, and (b) a composition comprising an inactive transpososome, under conditions suitable for transpososome activity, wherein a ratio of an amount of the inactive transpososome in the composition of (b) to an amount of the active transpososome in the composition of (a) determines the mean fragment size and a level of insertion bias.

Abstract: Compositions, systems, and methods for the display of analytes such as biomolecules are described. Display of analytes is achieved by coupling of the analytes to displaying molecules that are configured to associate with surfaces or interfaces. Arrays of analytes may be formed from the described systems for utilization in assays and other methods.

Abstract: The invention relates to a method of assembling a biosensor device comprising two or more biosensor units, wherein each unit comprises one or more biosensors comprising one or more carbon nanotubes (CNTs) coated with nucleic acid and one or more sensor molecules coupled to the nucleic acid, wherein each one of the one or more sensor molecules is capable of binding to a target molecule in a sample. Each biosensor unit is capable of detecting a different target molecule in a sample, and each unit comprises one or more biosensors each capable of detecting the same target molecule. The invention further relates to biosensor devices and methods for detecting target molecules in a sample using the same.

Abstract: Methods and compositions are provided for preparing DNA libraries. Enzymes, adaptors, and sample nucleic acids are provided in a single reaction mixture to facilitate library preparation.

Abstract: The disclosed embodiments relate to method, apparatus and system for high throughput single-cell DNA sequencing with droplet microfluidic. In an exemplary embodiment, a method for analyzing nucleic acids within a cell includes the steps of: (a) flowing individual cells together with a material capable of forming a polymer or microsphere that retains nucleic acids into a carrier fluid such that droplets are formed; (b) breaking the emulsion and collecting the microsphere hydrogels in an aqueous fluid; and (c) performing combinatorial labeling on the nucleic acids contained within the microspheres/hydrogels.

Abstract: Dysregulated expression of microRNAs (miRNAs) has emerged as a hallmark feature in human cancers. Aspects of the disclosure relate to methods for selecting optimal therapy for a patient from several alternative treatment options. A major clinical challenge in cancer treatment is to identify the subset of patients who will benefit from a therapeutic regimen, both in metastatic and adjuvant settings. The number of anti-cancer drugs and multi-drug combinations has increased substantially in the past decade, however, treatments continue to be applied empirically using a trial-and-error approach. Here methods and compositions are provided to determine the optimal treatment option for gastric cancer patients.

Abstract: A system and method for assaying high and low abundant biomolecules within a biological fluid sample is provided. The method includes: a) placing a biological fluid sample in contact with a first nanostructure surface; b) interrogating the sample with a light source, the sample in contact with the first nanostructure surface, the interrogation using a SERS technique; c) detecting an enhanced Raman scattering from at least one high abundant biomolecule type and producing first signals representative thereof; d) placing the sample in contact with a second nanostructure surface having a targeting agent that targets a low abundant biomolecule; e) interrogating the sample with the light source using the SERS technique; f) detecting the enhanced Raman scattering from the low abundant biomolecules and producing second signals representative thereof; and g) assaying the biological fluid sample using the first signals and the second signals.

Abstract: An example of a method includes providing a substrate with an exposed surface comprising a first chemical group, wherein the providing optionally comprises modifying the exposed surface of the substrate to incorporate the first chemical group; reacting the first chemical group with a first reactive group of a functionalized polymer molecule to form a functionalized polymer coating layer covalently bound to the exposed surface of the substrate; grafting a primer to the functionalized polymer coating layer by reacting the primer with a second reactive group of the functionalized polymer coating layer; and forming a water-soluble protective coating on the primer and the functionalized polymer coating layer. Examples of flow cells incorporating examples of the water-soluble protective coating are also disclosed herein.

Abstract: Embodiments of the present invention relate to analyzing components of a cell. In some embodiments, the present invention relate to analyzing components of a single cell. In some embodiments, the methods and compositions relate to sequencing nucleic acids. In some embodiments, the methods and compositions relate to identifying and/or quantitating nucleic acid, proteins, organelles, and/or cellular metabolites.

Abstract: Provided herein is a method for sequence analysis that comprises analyzing PCR reactions that each contain different portions of the same sample, wherein at least some of the primer pairs are in more than one PCR reaction and at least one of the PCR reactions contains some but not all of the primer pairs of the other reaction(s).

Abstract: Described herein is a method of sequencing a template that comprises a direct repeat, comprising: (a) in the same reaction, hybridizing a primer to a first site that is upstream of the first repeat sequence and hybridizing a primer to a second site that is upstream of the second repeat sequence, wherein the first and second sites are: (i) upstream of the first and second repeat sequences, respectively, and (ii) equidistant from the first and second repeat sequences; and (b) subjecting the hybridization product of (a) to a sequencing-by-synthesis sequencing reaction to produce a sequence read that comprises a combination of the first and second repeat sequences.

Abstract: This disclosure is related to a method of sequencing a single-stranded DNA using deoxynucleotide polyphosphate analogues and translocation of tags from incorporated deoxynucleotide polyphosphate analogues through a nanopore.

Abstract: The present invention is directed to methods, compositions and systems for analyzing sequence information while retaining structural and molecular context of that sequence information.

Abstract: The present invention concerns a novel method for the modification and/or custom-made design of artificial non-ribosomal peptide synthetases (NRPSs) from naturally available NRPSs. The artificial NRPSs are of predetermined length and amino acid composition and sequence. Via fusion of well-defined NRPS units (so-called “exchange units”) in a certain manner, using a specific sequence motif in the linker areas it is possible to construct artificial and/or modified NRPS assembly lines, which have the ability of synthesizing peptides of a desired structure.

Abstract: Provided herein are methods, compositions, and systems for multiplexed analysis of individual cells or cell populations. Cells encapsulated in beads and/or biomolecules are sequentially co-partitioned, allowing for analysis of two different types of biomolecules (e.g., RNA and DNA). The present invention leverages different polymer dissociation mechanisms, accompanied with barcoding of biomolecules (e.g., nucleic acid molecules) for multiplexed measurements in single cells. Sequential co-partitioning and barcode technology enables identification and quantitation of DNA and RNA from single cells.

Abstract: The present disclosure relates to use of a lectin receptor-like kinase LecRK-IX.1 as a protein for regulating insect resistance of Arabidopsis thaliana. A. thaliana with high resistance to Bemisia tabaci can be cultivated by reducing the expression of, or knocking out, an encoding gene of the protein. Therefore, Arabidopsis thaliana with the high-level resistance to Bemisia tabaci can be cultivated. The gene and its encoded protein can be applied to plant genetic improvement.

Abstract: Methods are disclosed for identifying antibacterial compounds which inhibit propagation of selected spectrum bacteria, which bacteria use specific tRNA to code for Ala, Met, Ser, or Leu that other bacteria do not use. In one embodiment, the selected spectrum bacteria use GCA to code for Ala, whereas other bacteria use a different codon to code for alanine. The methods involve determining whether putative inhibitors promote or inhibit complex formation between the tRNA and a bacterial ribosome, or between the tRNA and an aminoacyl synthetase. Compounds which promote or inhibit complex formation can disrupt protein production, which bacteria need to propagate. The identified antibacterial compounds can selectively inhibit bacterial propagation. By limiting their effects to the selected spectrum bacteria, these compounds can treat or prevent specific bacterial infections without disrupting the normal bacterial flora, the patients' microbiome, or causing antibacterial resistance.

Abstract: Methods, compositions, reaction mixtures, kits, and/or systems for producing a complementary sequence to a region in a target polynucleotide in a sample are provided. In some aspects, the methods, compositions, reaction mixtures, kits, and/or systems comprise subjecting the sample to a nucleic acid amplification reaction in a reaction mixture under conditions to yield the complete sequence to the region of the target polynucleotide. In some aspects, the complementary sequence produced is amplified.

Abstract: Bistable devices are constructed using a polynucleotide platform for sensing molecular events such as binding or conformational changes of target molecules. Uses include measurement of target concentration, measuring the effect of environmental condition (such as heat, light, or pH) on the target, or screening a library for molecules that bind the target or modulate its biological function. Devices comprise three regions: a top lid, bottom lid, and flexible linker or hinge between them. A device has an open configuration in which the top and bottom lid are separated, and a closed configuration they are bound close together. Binding domains or variations of the target molecule are fixed to a device so that when the molecular event occurs, the device switches from open to closed, or vice versa, which generates a signal. Optimal device design is determined by the signal modality (optical or electronic) used to measure closure of surface-immobilized devices.

Abstract: Systems, methods and kits are described for culturing one or more biological cells in a microfluidic device, including provision of nutrients and gaseous components configured to enhance cell growth, viability, portability, or any combination thereof. In some embodiments, culturing a single cell may produce a clonal population in the microfluidic device.

Abstract: Labeled nucleotide analogs comprising at least one avidin protein, at least one dye-labeled compound, and at least one nucleotide compound are provided. The analogs are useful in various fluorescence-based analytical methods, including the analysis of highly multiplexed optical reactions in large numbers at high densities, such as single molecule real time nucleic acid sequencing reactions. The analogs are detectable with high sensitivity at desirable wavelengths. They contain structural components that modulate the interactions of the analogs with DNA polymerase, thus decreasing photodamage and improving the kinetic and other properties of the analogs in sequencing reactions. Also provided are nucleotide and dye-labeled compounds of the subject analogs, as well as intermediates useful in the preparation of the compounds and analogs. Compositions comprising the compounds, methods of synthesis of the intermediates, compounds, and analogs, and mutant DNA polymerases are also provided.

Abstract: Methods and compositions for digital profiling of nucleic acid sequences present in a sample are provided.

Abstract: A multi-level microfluidic device is provided. The device includes a silicon wafer substrate and a stack of layers arranged on the silicon wafer substrate. The stack comprises a plurality of fluidic silicon layers, wherein each fluidic silicon layer includes a microfluidic structure at least one intermediate layer. The at least one intermediate layer is arranged between two fluidic silicon layers, and a fluid inlet and a fluid outlet in fluid connection with at least one of the fluidic silicon layers. Each layer in the stack is formed by deposition or growth. Methods for manufacturing microfluidic devices is also provided.

Abstract: Embodiments may include a method of determining a nucleic acid sequence. The nucleic acid may be DNA. The method may include forming, by a polymerase, a double-stranded nucleic acid molecule from a first nucleic acid strand and a second nucleic acid strand. Forming the double-stranded nucleic acid molecule may include adding a first compound to the second nucleic acid strand. The first compound may include a nucleotide attached to a third nucleic acid strand. The third nucleic acid strand may be attached to a moiety. The method may also include detaching the nucleotide from the third nucleic acid strand and the moiety. The method may further include measuring a change in an electrical characteristic of a transistor resulting from the moiety. Additionally, the method may include identifying the nucleotide based on the measured change in the electrical characteristic.

Abstract: CRISPR/Cas Systems are provided where a tuned guide RNA is used to discriminate between two protospacer sequences of same length that differ by one nucleotide.

Abstract: The present invention provides cell lines for high efficiency genome editing using cas/CRISPR systems, methods of generating such cells lines, and methods of generating mutations in the genome of an organism using such cell lines.

Abstract: A method of preparing a sample may include depositing an aqueous solution comprising copies of a primer into a layer of hydrophobic liquid on a substrate with a thermal inkjet device. A sample may include: a substrate; a layer of hydrophobic liquid on the substrate, the layer of hydrophobic liquid comprising a plurality of droplets of aqueous solution distributed in the layer, wherein the plurality of droplets contain: primers; a polymerase enzyme; deoxynucleotide triphosphates (dNTPs); and a target sequence for replication; and a cover, the cover contacting and covering the layer of hydrophobic liquid.

Abstract: Methods and compositions for digital profiling of nucleic acid sequences present in a sample are provided.

Abstract: Provided herein is a hierarchical superhydrophobic surface comprising an array of first geometrical features disposed on a substrate comprising a first material, and an array of second geometrical features disposed on the first features to form a hierarchical structure and a terminal level disposed on the second features, wherein the terminal level comprises a second material, the second material being different from the first material. The second material has a hydrophilicity different from the hydrophilicity of at least one of 1) the hydrophilicity of the second material and 2) hydrophilicity induced by the hierarchical structure. The present disclosure further methods of preparing hierarchical superhydrophobic surfaces and medical devices comprising the hierarchical superhydrophobic surfaces.

Abstract: A luminescence-enhancement system can include a luminescence-enhancement sensor having a substrate and a group of nanofingers with individual nanofingers that can be flexible and include a support end attached to the substrate, a distal tip positioned distally with respect to the substrate, and a middle portion between the support end and the distal tip. A coating of a metal or metal alloy can be applied to the substrate and the distal tip that is conductively continuous. The middle portion can be devoid of the coating or the coating at the middle portion is conductively discontinuous. A liquid ejector can be present and can include a jetting nozzle to eject a liquid droplet having a volume of 2 pL to 10 ?L. The group of nanofingers and the jetting nozzle can be positionable relative to one another for the droplet to be deposited on the group of nanofingers.

Abstract: A microfluidic emulsion droplet generation system and methods of use thereof are provided. The system may include a microfluidic substrate having a flow path configured and arranged for emulsion droplet generation, at least one textured surface in the flow path configured and arranged for inducing surface-mediated coalescence of emulsion droplets; and at least one channel junction in the flow path for emulsion droplet formation.

Abstract: In some embodiments, the disclosure relates generally to methods, as well as related compositions and kits for recombinase-mediated nucleic acid amplification, such as recombinase-polymerase amplification (RPA), of a nucleic acid template using at least one blocked primer that contains a 5? domain, at least one nucleotide that is cleavable by an RNase H enzyme, a 3? domain, wherein the primer is not extendable by a polymerase, and wherein the 3? domain has a length of 7-100 nucleotides, for example 10-30 nucleotides. These methods and the use of a blocked primer reduce or eliminate non-specific amplification products, such as primer dimers, which are generated in RPA reactions.

Abstract: Light detection devices and corresponding methods are provided. The devices include a reaction structure to contain a reaction solution and at least one reaction site that generates light emissions in response to incident excitation light after treatment with the reaction solution. The devices also include a plurality of light sensors and device circuitry. The devices further include a plurality of light guides extending toward at least one corresponding light sensor from input regions that receive the excitation light and the light emissions from at least one corresponding reaction recess. The light guides comprise a first filter region that filters the excitation light and permits the light emissions of a first wavelength to pass to the at least one corresponding light sensor, and a second filter region that filters the excitation light and the permits light emissions of a second wavelength to pass to the at least one corresponding light sensor.

Abstract: Provided are methods and systems useful for screening large libraries of effector molecules. Such methods and systems are particularly useful in microfluidic systems and devices. The methods and systems provided herein utilize encoded effectors to screen large libraries of effectors.

Abstract: A material with a nanotexture comprising structures extending from a substrate. The structures are modified by coating the nanotexture with a protective coating and partially removing the coating, exposing a portion of the structure for functionalization.

Abstract: This disclosure provides kits and methods for determining whether a subject is suffering from a liver disease, and/or whether the subject will be a responsive to a therapy for a liver disease. The disclosed methods comprise determining the miR profile of exosomes isolated from a body fluid of the subject.

Abstract: The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing and analyte characterization. Such polynucleotide processing may be useful for a variety of applications, including analyte characterization by polynucleotide sequencing. The compositions, methods, systems, and devices disclosed herein generally describe barcoded oligonucleotides, which can be bound to a bead, such as a gel bead, useful for characterizing one or more analytes including, for example, protein (e.g., cell surface or intracellular proteins), genomic DNA, and RNA (e.g., mRNA or CRISPR guide RNAs). Also described herein, are barcoded labelling agents and oligonucleotide molecules useful for “tagging” analytes for characterization.

Abstract: The present invention discloses a method for rapidly constructing amplicon library including the following steps: 1. Synthesizing a primer combination for constructing an amplicon library of a DNA sample, the primer combination of the amplicon library used to construct the DNA sample includes: a forward fusion primer designed according to the target amplicon, a reverse fusion primer designed according to the target amplicon, a forward universal primer and a reverse universal primer; 2. Constructing a PCR reaction system for the DNA sample; 3. Performing PCR. The method according to the present invention can be used to construct an amplicon library in a simple and rapid manner, and since a barcode is introduced before the start of PCR, the possibility of cross-contamination between the sample and the library is greatly reduced.

Abstract: In one aspect, the present invention relates to a method for obtaining structural information about an encoded molecule. The encoded molecule may be produced by a reaction of a plurality of chemical entities and may be capable of being connected to an identifier oligonucleotide containing codons informative of the identity of the chemical entities which have participated in the formation of the encoded molecule. In a certain embodiment, primers are designed complementary to the codons appearing on the identifier oligonucleotide, and the presence, absence or relative abundance of a codon is evaluated by mixing a primer with the identifier oligonucleotide in the presence of a polymerase and substrate (deoxy)ribonucleotide triphosphates and measuring the extension reaction. In another aspect, the invention provides a method for selecting compounds which binds to a target.

Abstract: A sample-reagent mixture is thermal cycled through a plurality of cycles. Each thermal cycle includes actuating a heater to heat the sample-reagent mixtures; and dispensing a fluid onto the sample-reagent mixture to cool the sample reagent mixture.

Abstract: Disclosed are catalytically active water-insoluble protein aggregates comprising fusion proteins which comprise a coiled-coil domain and a catalytic domain, methods of manufacturing such protein aggregates, and their use.

Abstract: The present invention generally relates to microfluidics and labeled nucleic acids. For example, certain aspects are generally directed to systems and methods for labeling nucleic acids within microfluidic droplets. In one set of embodiments, the nucleic acids may include “barcodes” or unique sequences that can be used to distinguish nucleic acids in a droplet from those in another droplet, for instance, even after the nucleic acids are pooled together. In some cases, the unique sequences may be incorporated into individual droplets using particles and attached to nucleic acids contained within the droplets (for example, released from lysed cells). In some cases, the barcodes may be used to distinguish tens, hundreds, or even thousands of nucleic acids, e.g., arising from different cells or other sources.

Abstract: The present disclosure relates to an organic electroluminescence device with enhanced out-coupling efficiency. The electroluminescence device comprises a substrate, a first electrode disposed on the substrate, an organic layer and a second electrode. The organic layer is formed between the first electrode and the second electrode. The organic layer comprises one or more nanoparticles forming protrusion features on the organic layer thereby forming a topographical structured organic layer in the electroluminescence device. The electroluminescence device is capable of reducing the surface plasmon polariton loss thereby, enhancing the light out-coupling the electroluminescence device.

Abstract: A system for analyzing a sample includes a chromatographic device, an electrospray source, and a mass resolving device. The chromatographic device is configured to separate components of the sample as a function of retention time within a chromatographic column. The electrospray source is configured to direct a first portion of a flow from the chromatographic device via a waste outlet to a pressurized waste reservoir, direct a second portion of the flow to an electrospray ionization outlet to form a spray, and charge and desolvate the spray to form ions of the components of the sample. A flow rate of the second portion of the liquid flow is substantially determined by a pressure of the pressurized waste reservoir. The mass resolving device configured to receive the ions and characterize the mass-to-charge ratio of the ions.

Abstract: The present disclosure relates to compounds comprising a negatively-charged polymer moiety which is capable of entering a nanopore and upon entering a nanopore in the presence of positive ions results in an increased flow of the positive ions through the nanopore. The present disclosure provides methods of preparing the compounds and for their use as nanopore-detectable tags, in particular, for nanopore-based nucleic acid detection and sequencing.

Abstract: An apparatus suitable for single molecule sequencing. The apparatus includes at least one nanowell, a plurality of nucleic acid immobilization moieties, and a plurality of types of nucleic acid fragments. The nanowell has an observation zone. The nucleic acid immobilization moieties are disposed in or proximate to the observation zone. The nucleic acid fragments are immobilized to the nucleic acid immobilization moieties, respectively. At least one polymerase is disposed in the observation zone. A method of sequencing nucleic acid molecules using the above-mentioned apparatus is provided.

Abstract: The present disclosure provides methods, compositions, kits and systems for nucleic acid amplification. In some embodiments, nucleic acid amplification methods include subjecting the nucleic acid to be amplified to partially denaturing conditions. In some embodiments, nucleic acid amplification methods include amplifying without fully denaturing the nucleic acid that is amplified. In some embodiments, the nucleic acid amplification method employs an enzyme that catalyzes homologous recombination and a polymerase. In some embodiments, methods for nucleic acid amplification can be conducted in a single reaction vessel and/or in a single continuous liquid phase of a reaction mixture, without need for compartmentalization of the reaction mixture or immobilization of reaction components.

Abstract: The present invention is concerned with compositions and methods for improving the rate of correct sample identification in indexed nucleic acid library preparations for multiplex next generation sequencing by exonuclease treatment after protective adapters are ligated to target polynucleotides to degrade unincorporated adapters prior to amplification and sequencing.

Abstract: Provided are a vesicular adaptor and a single-chain cyclic library constructed by using the adaptor. The library can be used for RNA sequencing and other sequencing platforms dependent on a single-stranded cyclic library, and has the advantage of high throughput sequencing, high accuracy and simple operations.