Abstract: Described herein are methods, compositions, kits, and uses thereof for analysis of nucleic acid segments comprising a repeating A/T-rich segment, wherein the repeating A/T-rich segment is: (i) a homopolymeric segment comprising at least 10 A residues, at least 10 T residues, or at least 10 U residues, wherein the at least 10 A, T, or U residues are consecutive or interrupted once by one to three other nucleotides; or (ii) a segment comprising (TnA)m, (ATn)m, (TAn)m, or (AnT)m, wherein n is 2 or greater and m is such that the length of the repeating A/T-rich segment is 10 or more residues.

Abstract: Methods, devices, and systems for performing intermittent detection during analytical reactions are provided. Such methods facilitate collection of reaction data from disparate reaction times. Further, such methods are useful for reducing photo-induced damage of one or more reactants in an illuminated analytical reaction at a given reaction time. In preferred embodiments, the reaction mixture is subjected to at least one illuminated and non-illuminated period and allowed to proceed such that the time in which the reaction mixture is illuminated is less than a photo-induced damage threshold period.

Abstract: Systems and methods for storing large data sets, such as genetic sequence information. Within a “targeted subset” of positions with information, the system stores, both variant states and missing states at each position. Reference states are not stored, but are inferred within the targeted subset when neither a variant nor a missing state is stored at a given position. The absence of a variant state at a given position is assumed to be a reference state. The criteria for missing data are defined in pre-processing and are customizable based on the use case. For example, each data point may represent the genetic information of a sample at a position in the genome. The targeted subset may represent those positions that were included in a sequencing test.

Abstract: The present disclosure provides for methods and compositions useful for imaging inflammation and inflammatory disease markers with an affinity for TREM-1 antibodies. The methods and compositions can include a labeled probe having a TREM-1 antibody and a radiolabel.

Abstract: Methods, compositions and kits for capturing, detecting and quantifying mature small RNAs are provided herein. Embodiments of the methods comprise ligating 5? and 3? ligation adaptors to the 5? and 3? ends of the mature small RNAs, respectively, in the presence of 5? and 3? semi-degenerate ligation splints to generate a ligation product. Other embodiments comprise reverse transcribing polyadenylated mature small RNA with a universal reverse transcription primer and ligating an adaptor to the 3? end of the cDNA in the presence of a semi-degenerate ligation splint to generate a cDNA ligation product.

Abstract: A kit for use with a nucleic acid sequencing instrument can include a plurality of combinatorial barcodes sequences meeting the following criteria: each of the combinatorial barcode sequences comprise a plurality of iterations of a sequence motif, where the sequence motif comprises a first nucleotide base from a first group of nucleotide bases followed by a second nucleotide base from a second group of nucleotide bases, the first group and the second group differing from each other; and the plurality of combinatorial barcode sequences is at least 1,000,000 different barcode sequences.

Abstract: The present invention is directed to methods for identifying the presence of one or more target nucleotide sequences in a sample that involve a ligation and/or polymerase reaction. In some embodiments, the ligation products formed in the ligation process of the present invention are subsequently amplified using a polymerase chain reaction. The ligated product sequences or extension products thereof are detected, and the presence of one or more target nucleotide sequences in the sample is identified based on the detection.

Abstract: The present disclosure provides a HTP microbial genomic engineering platform that is computationally driven and integrates molecular biology, automation, and advanced machine learning protocols. This integrative platform utilizes a suite of HTP molecular tool sets to create HTP genetic design libraries, which are derived from, inter alia, scientific insight and iterative pattern recognition. The HTP genomic engineering platform described herein is microbial strain host agnostic and therefore can be implemented across taxa. Furthermore, the disclosed platform can be implemented to modulate or improve any microbial host parameter of interest.

Abstract: A biosensor platform apparatus and method are provided that can detect, purify and identify nucleic acid (DNA and RNA) biomarkers in complex biological fluids. The methods use a two-stage molecular based approach. The first stage screens for specific nucleic acid-based biomarkers in complex biological fluids by electrochemical detection of DNA:RNA hybridization and facilitates the removal of remaining complex media constituents. The first stage utilizes probes within a tunable nanoporous electrode. The second stage identifies the purified specific hybrids by single-molecule conductance measurements via break junction scanning. Identification can be assisted with a library of conductance measurements. The methods can provide strain level information that can be used for identifying anti-microbial resistance in detected pathogens. Collection of RNA targets allows for biomarker detection and identification without the need for amplification and can provide information about the viability of the sample organism.

Abstract: Flow cell devices, cartridges, and systems are described that provide reduced manufacturing complexity, lowered consumable costs, and flexible system throughput for nucleic acid sequencing and other chemical or biological analysis applications. The flow cell device can include a capillary flow cell device or a microfluidic flow cell device.

Abstract: A microfluidic thermalization chip, a system using such a chip and a PCR method for detecting DNA sequences. The chip contains a block of material in which a cavity is configured to contain at least one fluid. The cavity includes at least one inlet orifice and at least one outlet orifice. The inlet orifice for the fluid is connected to at least one, preferably at least two, fluid-injecting channels. Further, the chip includes at least one microfluidic channel for bypassing the cavity. The channel is connected by a first end to at least one of the fluid-injecting channels. The junction between the bypassing channel and the fluid-injecting channel is located at a distance L from the inlet orifice of the fluid-injecting channel. The distance L is preferably smaller than 2 cm.

Abstract: The invention includes methods for determining the presence of a latent viral population by analyzing an RNA population from the virus with digital techniques, such as digital PCR or by sequencing cDNA produced from the RNA. The invention additional includes methods for determining the presence of latent viral populations by detecting and/or quantifying enzymes that are uniquely associated with the virus, e.g., reverse transcriptases.

Abstract: A method of preparing a library of small interfering RNA (siRNA) expression systems for producing siRNA for silencing of target genes by inducing degradation of target gene RNA expression products includes: (i) isolating RNA of one or more target genes from a cell population; (ii) generating RNA fragments from the isolated RNA; (iii) converting the RNA fragments into dsDNA fragments; and (iv) cloning the dsDNA fragments into vectors for forming cloned vectors, each vector including one or more promoters and at least one restriction enzyme site capable of accepting the insertion of at least one dsDNA fragment such that siRNA can be produced. Methods for producing siRNA from the siRNA expression system and methods of identifying a functional target gene for treatment by using the siRNA produced from the siRNA expression system and for identifying RNAi therapeutics are also provided.

Abstract: The present invention relates to a method and a composition for detecting a target nucleic acid sequence using a cleaved complementary tag fragment. Specifically, the present invention relates to a method for linking a complementary tag sequence to a PCR primer so that a tagging can be produced by a restriction enzyme during a PCR reaction, diversifying the complementary tag sequence to be linked to each primer by utilizing factors such as length and nucleic acid combination, etc., and distinguishing the target sequence using the same.

Abstract: The present invention relates to utilizing terminal erythroid differentiation (TED) as a biomarker for prognosis and as a therapeutic target in myeloid malignancies, in particular myelodyplastic syndromes. The present invention relates to identifying patients with myelodysplastic syndromes at risk for poor survival/outcomes who would benefit from aggressive treatment, by characterizing their TED profile using protein and gene expression markers and combinations thereof.

Abstract: Melanophilin (MLPH) of the present invention is involved in differentiation into an adipocyte or fat accumulation, and accordingly, obesity can be treated or prevented by inhibiting the MLPH. Further, by measuring an expression level of the MLPH, obesity can be diagnosed and treated, and therapeutic agents for obesity and agents regulating differentiation into adipocytes can be screened.

Abstract: FRET-labeled compounds are provided for use in analytical reactions. In certain embodiments, FRET-labeled nucleotide analogs are used in place of naturally occurring nucleoside triphosphates or other analogs in analytical reactions comprising nucleic acids, for example, template-directed nucleic acid synthesis, DNA sequencing, RNA sequencing, single-base identification, hybridization, binding assays, and other analytical reactions.

Abstract: An object is to provide a component for molecular Computing and a method of molecular Computing. A component for molecular Computing, the component comprising: a microsphere including pores, at least some of which are open on a surface of the microsphere, and a plurality of modules grafted on the microsphere wherein each of the modules is a continuous séquence of nucleic acid base.

Abstract: Disclosed are methods for determining the immunological status of the adaptive immune system of a subject by identifying and quantifying rearranged DNA (and/or subsequently transcribed RNA) sequences encoding T cell receptor (TCR) and/or immunoglobulin (IG) polypeptides, in a lymphoid DNA-containing sample from the subject. TCR and/or IG sequence diversity and sequence distribution permit immunocompetence and immune repertoire assessment and reflect the degree of T cell or B cell clonality and clonal expansion in the sample. Methods for stratifying patient populations on the basis of immunocompetence including likelihood of responding to immunotherapy are also described.

Abstract: The subject disclosure presents systems and methods for improved meso-dissection of biological specimens and tissue slides including importing one or more reference slides with annotations, using inter-marker registration algorithms to automatically map the annotations to an image of a milling slide, and dissecting the annotated tissue from the selected regions in the milling slide for analysis, while concurrently tracking the data and analysis using unique identifiers such as bar codes.

Abstract: A method for identifying a nucleic acid template that includes (a) providing a plurality of primer-template hybrids, wherein a first hybrid of the plurality includes a first template hybridized to a first primer, and wherein a second hybrid of the plurality includes a second template hybridized to a second primer, the second primer having a ternary complex inhibitor moiety at the 3? end; (b) delivering polymerases and nucleotides to the plurality, whereby the first hybrid binds a polymerase and nucleotide to form a stabilized ternary complex and whereby the second hybrid does not bind a polymerase and nucleotide to form a stabilized ternary complex; and (c) detecting the stabilized ternary complex to identify the first template.

Abstract: Devices and methods are provided for collecting and handling difficult sample types.

Abstract: Methods and systems and related compositions for separating through a solid matrix a mixture comprising a nucleic acid together with a target compound having a water solubility equal to or greater than 0.01 mg per 100 mL, which can be used for managing fluid flow, biochemical reactions and purification of the nucleic acid or other target analytes.

Abstract: This invention allows ultra-low levels of virtually any biological analyte to be detected and quantified rapidly, simply and inexpensively with an electrochemical biosensor using a novel electrochemically detectable tag that replaces optical labels. The tag binds to an analyte directly, or indirectly using one or more ligands and particles, and consists of a quadruplex electrochemically detectable oligonucleotide rich in guanine, or a single-stranded electrochemically detectable oligonucleotide rich in guanine that self-assembles into a quadruplex electrochemically detectable oligonucleotide when exposed to cations that enable quadruplex self-assembly. Quadruplex electrochemically detectable oligonucleotide tags are exposed, adsorbed or hybridized at the surface of a biosensor working electrode. An electrochemical technique facilitates the quadruplex tags to produce 8-oxoguanine oxidation signals proportional to the analyte level in the samples.

Abstract: Methods and uses for molecular tags are disclosed. Molecular tags may be attached to nucleic acid molecules. The attachment of the nucleic acid molecules prior to PCR amplification and sequencing improves the accuracy of genetic analysis and detection of genetic variations and diversity. Molecular tags may also be used for detection of drug-resistant variants. Methods for using molecular tags for determining and correcting PCR errors and/or sequencing error are also disclosed.

Abstract: This disclosure relates to compositions and methods for single-step, multi-stage amplification reactions that combine many stages of sample preparation process in a single tube reaction. The disclosed technology provides a mean of performing multiplexed nested PCR in a single vessel, without any need of purification steps, and is based on the use of three sets of primers: a pair of outer primers, a pair of inner primers that are nested within the pair of outer primers, and tail primers that are complementary to tails on the inner primers. By adjusting the temperature conditions, annealing temperatures of the primers, number of amplification cycles, and the concentrations of the outer, inner, and tail primers, it is possible to carry out multiplexed nested PCR in a single vessel.

Abstract: The present disclosure provides compositions, methods and kits for Omega amplification technologies. In addition, the present disclosure provides compositions, methods and kits for universal FQ probe and for G-quadruplex detection methods for use in isothermal amplification technologies.

Abstract: The present disclosure relates to kits for determining the length of a region of tandem repeats in a subject's genome. In some embodiments, the region of tandem repeats in telomeres.

Abstract: Disclosed herein are methods and compositions for determining the copy number of a first target nucleic acid as compared to the copy number of a second target nucleic acid in a single well with a single detection label. For example, disclosed herein are methods and compositions for determining the copy number of a first target nucleic acid as compared to the copy number of a second target nucleic acid by a monochrome multiplex quantitative PCR (MMQPCR) in a single well with a single detection label.

Abstract: A nucleic acid patch method for amplifying target nucleic acid sequences in circulating free DNA or residual DNA samples where the defining ends of the target nucleic acid sequences are unknown.

Abstract: Methods for performing multiplex PCR-based enrichment of a target substrate are provided. Systems and methods for generating a sequencing library are also provided.

Abstract: A computer-implemented method according to one embodiment includes identifying principal components for a dataset defined by data instances and features corresponding to the data instances, identifying, for at least one of the data instances, at least some of the principal components, wherein the identified principal components are determined to be salient for said at least one data instance, and determining, for said at least one of the data instances, one or more salient features corresponding to the identified salient principal components.

Abstract: Methods are provided for a rapid, low cost approach to monitoring an amplification reaction. This includes monitoring the progress of isothermal or PCR amplification reactions to completion using pH-sensitive dyes that are either colored or fluorescent. Compositions are described that include a mixture of a DNA polymerase, deoxyribonucleotide triphosphate and Tris buffer in the range of 1.5 mM Tris to 5 mM Tris or equivalent.

Abstract: Next Generation DNA sequencing promises to revolutionize clinical medicine and basic research. However, while this technology has the capacity to generate hundreds of billions of nucleotides of DNA sequence in a single experiment, the error rate of approximately 1% results in hundreds of millions of sequencing mistakes. These scattered errors can be tolerated in some applications but become extremely problematic when “deep sequencing” genetically heterogeneous mixtures, such as tumors or mixed microbial populations. To overcome limitations in sequencing accuracy, a method Duplex Consensus Sequencing (DCS) is provided. This approach greatly reduces errors by independently tagging and sequencing each of the two strands of a DNA duplex. As the two strands are complementary, true mutations are found at the same position in both strands. In contrast, PCR or sequencing errors will result in errors in only one strand.

Abstract: The present invention relates to a combination product for detecting a target marker simply and with high sensitivity. More specifically, the present invention relates to a combination product for detecting a target marker in a biological sample in combination with a target marker binding molecule which is capable of binding specifically to the target marker in the biological sample, the combination comprising, at least: (a) a first binding agent comprising a first binding molecule which is capable of directly or indirectly binding specifically to the target marker binding molecule, and a labeling substance; (b) a linker molecule which is capable of binding specifically to the first binding agent; and (c) a second binding agent which is capable of binding specifically to the linker molecule, and comprises a second binding molecule and a labeling substance.

Abstract: One aspect of this disclosure relates to a computer-implemented method for determining a force acting on at least part of a structure, for example a biological structure, such as a DNA molecule. The method comprises controlling a light-sensitive system, e.g. of a microscope, to determine light information based on light from the structure. The light is incident on at least a part of the light sensitive system. The light-sensitive system may be said to capture the light from the structure. The at least part of the structure comprises one or more optically active entities, such as DNA intercalator molecules and donor/acceptor fluorophores. At least one of (i) an optical activity of the entities and (ii) a quantity of the entities depends on the force acting on the at least part of the structure. Furthermore, the light information defines a light property value associated with said at least part of the structure.

Abstract: Described herein are nanopore protein conjugates that can be used in DNA sequencing reactions. The nanopore protein conjugates includes a nanopore protein monomer that is joined to a DNA binding domain. The nanopore protein monomer is available to oligomerize with other nanopore protein monomers, while the DNA binding domain is available to bind to a template DNA strand. In certain examples, the nanopore protein monomer is an alpha-hemolysin monomer or variant thereof and the DNA binding domain is an Sso7d protein or variant thereof, such as an Sso7d-like protein. Also provided are nanopore protein assemblies incorporating the nanopore protein conjugates, along with methods of using the nanopore protein assemblies in sequencing reactions.

Abstract: The present invention is directed to methods for assaying for the presence of SARS-CoV-2 in a sample, including a clinical sample, and to oligonucleotides, reagents, and kits useful in such assays. In particular, the present invention is directed to such assays that are rapid, accurate and specific for the detection of SARS-CoV-2.

Abstract: The present invention generally relates to a method for analyzing and identification of the plurality of lipids in a sample that is profiled using a combined Liquid Chromatography-Mass Spectrometry (LC-MS) technique, comprising the steps of: a) providing a list of Liquid Chromatography-Mass Spectrometry (LC-MS)-based mass features; b) deconvoluting said list of LC-MS-based mass features; c) inferring daughter ions from the deconvoluted list of LC-MS-based mass features; d) identifying one or more parental exact masses from the inferred daughter ions; e) scoring each of the one more parental exact masses based on the inferred daughter ions; f) determining characteristic mass features in response to the scoring of each of the one or more parental exact masses; and g) determining each of the plurality of lipids based on the characteristic mass features thereof. In particular, the present invention also relates to identification of the plurality of lipids undergoing in-source fragmentation.

Abstract: Real time electronic sequencing devices, chips, and systems are described. Arrays of nanoFET devices are used to provide sequence information about a template nucleic acid in a polymerase-template complex bound to the nanoFET. The nanoFET devices typically have a source, a drain and a gate comprising a nanowire. A single polymerase enzyme complex comprising a polymerase enzyme complexed with the template nucleic acid is bound to the gate. The polymerase is bound to the gate non-covalently through a polymeric binding agent that has two strands, each strand interacting with the nanowire such that the polymerase is in a central location between the strands with the polymeric binding agent extending away from the polymerase complex along the nanowire in both directions.

Abstract: Disclosed are compositions and methods for the preparation of RNA libraries for sequencing, gene expression profiling, microarray and other uses and for simplification of the library preparation process. The disclosure provides blocking oligonucleotides which bind to byproduct nucleic acid molecules formed during the ligation of adapters to nucleic acid segments prior to sequencing and inhibit or block amplification of the byproduct nucleic acid molecules in subsequent amplification reactions. Methods for library preparation using blocking oligonucleotides are also provided.

Abstract: This application describes methods and compositions for detecting and treating vimentin-associated neoplasia. Differential methylation of the vimentin nucleotide sequences has been observed in vimentin-associated neoplasia such as neoplasia of the upper or lower gastrointestinal tract, pancreas, and/or bladder.

Abstract: This disclosure relates to a glucose-sensing electrode including a nanoporous metal layer and a maltose-blocking layer formed over the nanoporous metal layer. The nanoporous metal layer is capable of oxidizing both glucose and maltose without an enzyme specific to glucose or maltose in the glucose-sensing electrode. The maltose-blocking layer has porosity that permits glucose to pass therethrough and inhibits maltose from passing therethrough toward the nanoporous metal layer.

Abstract: The invention features a fully-integrated rapid molecular diagnostic device that is low-cost, easy to manufacture, and simple to use. The device can serve as a molecular diagnostic platform for any disease, requiring little or no preparation or customization and can be made from simple materials (e.g., paper and adhesive film), making it inexpensive, portable, and disposable. The invention also provides methods of using the device for detection of one or more targets in a sample.

Abstract: The invention includes methods and apparatus for separating mutations, especially rare and unknown mutations, using heteroduplex binding proteins. Nucleic acids may optionally be nicked at or near the mutation in order to promote heteroduplex binding protein recognition and binding. In particular, using the disclosed methods, it is possible to separate heteroduplexed nucleic acid strand pair from homoduplexed nucleic acid strand pairs having similar sequences and being at a much higher concentration. Once the heteroduplexed nucleic acids are isolated and recovered, it is straightforward to analyze the sequences of the heteroduplexed nucleic acids, e.g., using sequencing or hybrid assays.

Abstract: The invention provides compositions and methods for simultaneously determining the presence or absence of fetal aneuploidy and the relative amount of fetal nucleic acids in a sample obtained from a pregnant female. The method encompasses the use of sequencing technologies and exploits the occurrence of polymorphisms to provide a streamlined noninvasive process applicable to the practice of prenatal diagnostics.

Abstract: According to one embodiment of the present invention, a structure includes: a substrate having a patterned surface of one or more binding sites; and a molecular shape made by a polynucleotide platform having a shape corresponding to a shape of a binding site of the one or more binding sites, the molecular shape being bound to one of the one or more binding sites.

Abstract: Systems and methods are provided for assessing a sample obtained from a subject. The computerized method includes receiving, by receiving circuitry, a dataset associated with the sample, the dataset comprising quantitative expression data for LRRN3, CDKN1C, PALLD, SASH1, RGL1, and TNFRSF17. A processor generates, based on the received dataset, a score that is indicative of a predicted smoking status of the subject. The predicted smoking status may classify the subject as a current smoker or as a non-current smoker.

Abstract: A region of the Trichomonas vaginalis AP65-1 gene has been identified which is useful for performing amplification assays to determine specifically whether T. vaginalis is present in the sample being tested. Oligonucleotides useful for performing thermal Strand Displacement Assay (tSDA) reactions on this gene are disclosed. The disclosed oligonucleotides can be used in an assay which is specific for multiple strains of T. vaginalis and which does not show cross reactivity with the genomes of other microorganisms or with human DNA.

Abstract: Provided herein are methods and systems for biochemical analysis, including compositions and methods for processing and analysis of small cell populations and biological samples (e.g., a robotically controlled chip-based nanodroplet platform). In particular aspects, the methods described herein can reduce total processing volumes from conventional volumes to nanoliter volumes within a single reactor vessel (e.g., within a single droplet reactor) while minimizing losses, such as due to sample evaporation.