Abstract: Disclosed herein is a method of quantifying a mutant allele burden of a target gene in a subject. The method includes providing a first plasmid that includes a mutant allele sequence and an internal control sequence, and a second plasmid that includes a wild-type allele sequence and the internal control sequence, and subjecting DNA of the subject to quantitative polymerase chain reaction to measure a mutant allele expression level of the target gene, so as to determine the mutant allele burden of the target gene in the subject based on a standard curve of the mutant allele burden of the target gene created by serial dilution of the first and second plasmids.

Abstract: Provided herein are methods and compositions for synthesizing 5?Capped RNAs wherein the initiating capped oligonucleotide primers have the general form m7Gppp[N2?Ome]n[N]m wherein m7G is N7-methylated guanosine or any guanosine analog, N is any natural, modified or unnatural nucleoside, “n” can be any integer from 0 to 4 and “m” can be an integer from 1 to 9.

Abstract: The present disclosure features useful compositions and methods to treat nucleotide repeat expansion disorders, e.g., in a subject in need thereof. In some aspects, the compositions and methods described herein are useful in the treatment of disorders associated with MSH3 activity.

Abstract: Methods for identifying and optionally treating subjects, e.g., subjects who have Chronic Rhinosinusitis (CRS), based on the detection of elevated levels of soluble P-glycoprotein in nasal secretions. The methods of treatment can include administration of a therapeutically effective amount of a P-glycoprotein inhibitor.

Abstract: Proteases regulate a wide range of normal cellular functions where dysregulated activity is observed in various diseases. Compositions and methods use protease activity multiplexed bead-based immunoassays to profile protease activity. This platform technology integrates protease activity measurements with total protein quantification techniques. It represents a significant improvement over existing detection techniques by allowing for multiplexed, sensitive active protease measurements in complex biological samples. Exemplary multiplexed detections are realized in a single assay using a minute sample amount (e.g., 5 ?l) for active recombinant MMP-1, -2, -3, -7, 9, and 12 and those same MMPs in cell culture supernatant, menstrual fluid effluent, and peritoneal aspirates. This multiplexed platform achieves high level of sensitivities equal to or better than existing leading single-plex detection strategies.

Abstract: A multi-scale habitat information-based method and device for detecting and controlling water and fertilizer for crops in seedling stage: performing fusion analysis on multi-scale features of the water and fertilizer stress of crops on the basis of crop canopy-scale three-dimensional laser scanning information, foliage-scale polarization-hyperspectral imaging information, and micro-scale micro-CT scanning information; combining the real-time feedback of the temperature, humidity, illumination and substrate moisture content within a crop growing greenhouse; by means of multi-information fusion modeling, comprehensively determining and feeding back the water and fertilizer stress of the crops as well as water requirement and fertilizer requirement information, and providing policy information for the amount of fertilization and irrigation.

Abstract: The present invention relates to methods of joining two or more double-stranded (ds) or single-stranded (ss) DNA molecules of interest in vitro, wherein the distal region of the first DNA molecule and the proximal region of the second DNA molecule of each pair share a region of sequence identity. The method allows the joining of a large number of DNA fragments, in a predetermined order and orientation, without the use of restriction enzymes. It can be used, e.g., to join synthetically produced sub-fragments of a gene or genome of interest. Kits for performing the method are also disclosed. The methods of joining DNA molecules may be used to generate combinatorial libraries useful to generate, for example, optimal protein expression through codon optimization, gene optimization, and pathway optimization.

Abstract: The objective of the present invention is to provide a versatile method that makes highly sensitive detection of nucleic acid, and the like, possible even if a solution has a high salt concentration. This objective has been achieved through a method for detecting a target substance in a solution through EIS impedance measurement, wherein the solution includes at least the target substance, an impedance observation substance A, and a substance B for causing the charge mobility of the impedance observation substance to vary, and the substance B for causing the charge mobility to vary has a property of being taken into the target substance.

Abstract: This invention relates to methods, compositions and kits for extending a polynucleotide and for preparing sequencing library of polynucleotides involving generating modified target polynucleotide on an adaptor template oligonucleotide and tagging one or two strands of a target sequence. The sequencing library is suitable for massive parallel sequencing and comprises a plurality of double-stranded nucleic acid molecules.

Abstract: Methods and systems for digital measurements are provided. In an embodiment, the method includes producing a plurality of droplets, wherein at least one of the droplets of the plurality of droplets contains an analyte molecule from a sample; measuring at least a first portion of the plurality of droplets to determine individual volumes of droplets in the first portion of the plurality of droplets; analyzing at least a second portion of the plurality of droplets to determine a number of droplets in the second portion of the plurality of droplets that contain the analyte molecule; and using individual volumes of the droplets in the first portion of the plurality of droplets and the number of droplets in the second portion of the plurality of droplets that contain the analyte molecule to determine the concentration of the analyte molecule in the sample.

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 invention is generally related to systems and methods for producing a plurality of droplets. The droplets may contain varying species, e.g., for use as a library. In some cases, the fluidic droplets may be rigidified to form rigidified droplets (e.g., gel droplets). In certain embodiments, the droplets may undergo a phase change (e.g., from rigidified droplets to fluidized droplets), as discussed more herein. In some cases, a species may be added internally to a droplet by exposing the droplet to a fluid comprising a plurality of species.

Abstract: The invention provides compositions comprising nucleic acid complexes for use in screening compounds based on their ability to modulate binding interactions, wherein the compounds are barcoded.

Abstract: The present disclosure provides, among other things, methods for colorectal cancer screening and compositions related thereto. In various embodiments, the present disclosure provides methods for colorectal cancer screening that include analysis of methylation status of one or more methylation biomarkers, and compositions related thereto. In various embodiments, the present disclosure provides methods for colorectal cancer screening that include screening methylation status of one or more methylation biomarkers in cfDNA, e.g., in ctDNA. In various embodiments, the present disclosure provides methods for colorectal cancer screening that include screening methylation status of one or more methylation biomarkers in cfDNA, e.g., in ctDNA, using MSRE-qPCR.

Abstract: A system includes a memory and a processor configured to execute computer instructions stored in the memory that when executed cause the system to perform operations. The computer instructions include a learning component that includes one or more trained models to learn pathogenic features of a given fungal species based on learning from a plurality of stored fungal species images. A fungal identifier component employs the trained models to determine pathogenic parameters of an unidentified fungal species image based on the learned pathogenic features. The fungal identifier component generates an output file that classifies the unidentified fungal species image according to the determined pathogenic parameters from the unidentified fungal species image.

Abstract: Methods and compositions for high-throughput, single cell analyses are provided. The methods and compositions can be used for analysis of genomes and transcriptomes, as well as antibody discovery, HLA typing, haplotyping and drug discovery.

Abstract: The invention provides compositions and methods for making closed nucleic acid structures in which one or both strands are continuous. The closed nucleic acid structures can be used as sequencing templates among other applications.

Abstract: The present invention is directed to methods and compositions for long fragment read sequencing. The present invention encompasses methods and compositions for preparing long fragments of genomic DNA, for processing genomic DNA for long fragment read sequencing methods, as well as software and algorithms for processing and analyzing sequence data.

Abstract: The invention is directed to methods for determining antigen-specific T cells generally and to T cell receptors which bind an epitope of the Wilms' tumor antigen-1 (WT1) protein specifically. The disclosure also provides polynucleotides encoding the TCRs, engineered cells exogenously expressing the TCRs, and methods of making and using the TCRs and/or cells expressing the TCRs.

Abstract: The present disclosure relates to an in vitro method for releasing analytes from a sample involving contacting the sample with (i) a chaotropic agent, (ii) an organic solvent, (iii) a detergent, and (iv) at least one agent providing bicarbonate ions. The present disclosure further relates to a release agent for releasing analytes from a sample having the aforesaid compounds. Moreover, the present disclosure relates to uses, kits, methods and devices related to the method and the release agent of the present disclosure.

Abstract: The present invention relates to variant of Family A polymerases able to synthesize a nucleic acid fragment without template and to incorporate a reversible modified terminator nucleotide during the nucleic acid fragment synthesis. The present invention further relates to uses thereof for enzymatic synthesis of nucleic acid molecules.

Abstract: Compositions are provided for 3? adapters and methods of use are provided that include methods requiring a plurality of ligation steps involving a single-stranded target polynucleotide and 3? and 5? adapters. Embodiments of the 3? adapters comprise a cleavable linker positioned between a single-stranded region and a double-stranded region. Upon ligating the 3? adapters, the single-stranded region is released by cleaving the cleavable linker.

Abstract: A method of detecting the presence of a prostate cancer field defect in a human subject comprising the step of (a) obtaining genomic DNA from the human subject and (b) quantitating methylation in at least one target region selected from the group consisting of CAV1, EVX1, MCF2L, FGF1, NCR2 and WNT2 and EXT1 and SPAG4 target, wherein significant methylation changes indicate the presence of prostate cancer or a prostate cancer field defect, wherein the change is relative to tissue from a second human subject who does not have prostate cancer.

Abstract: Fluorescence imaging system designs are described that provide larger fields-of-view, increased spatial resolution, improved modulation transfer and image quality, higher spatial sampling frequency, faster transitions between image capture when repositioning the sample plane to capture a series of images (e.g., of different fields-of-view), and improved imaging system duty cycle, and thus enable higher throughput image acquisition and analysis for genomics and other imaging applications.

Abstract: The disclosure of the preset invention relates to a new method for preventing or treating tumor progression or tumor recurrence comprising administering to a subject in need thereof a therapeutically effective amount of an agent disrupting Musashi-1 (MSI1)/Argonaute 2 (AGO2) interaction. A composition or pharmaceutical composition for preventing or treating tumor progression or tumor recurrence is also provided, which comprises an agent disrupting Musashi-1 (MSI1)/Argonaute 2 (AGO2) interaction.

Abstract: This disclosure provides, among other things, a method for processing a membrane comprising rolling circle amplification (RCA) products. In some embodiments, this method may comprise: (a) obtaining a porous capillary membrane that comprises fluorescently labeled RCA products that are in or on the membrane; (b) depositing a curable polymer onto the membrane; and (c) curing the curable polymer to encapsulate the RCA products in a solid. In some embodiments, the curable polymer may be a silicone and may be transparent in its solid form. A kit for performing the method and a composition made by the method are also provided.

Abstract: Various methods, systems, computer readable media, and graphical user interfaces (GUIs) are presented and described that enable a subject, doctor, or user to characterize or classify various types of cancer precisely. Additionally, described herein are methods, systems, computer readable media, and GUIs that enable more effective specification of treatment and improved outcomes for patients with identified types of cancer. Some embodiments of the methods, systems, computer readable media, and GUIs described herein comprise obtaining RNA expression data and/or whole exome sequencing (WES) data for a biological sample from a subject; determining a molecular-functional (MF) profile for the subject; identifying an MF profile cluster with which to associate the MF profile for the subject; and clustering the plurality of MF profiles to obtain the MF profile clusters.

Abstract: The present invention provides, among other things, methods of quantitating mRNA capping efficiency, particularly for mRNA synthesized in vitro. In some embodiments, the methods comprise chromatographic methods of quantifying capping efficiency and methylation status of the caps.

Abstract: Molecules or salts thereof are provided, having the structure in Formula I, wherein n2 and n4 are the same or different and are independently 1, 2, or 3, and n3 is 1 to 20; X is oxygen, nitrogen, or sulfur; wherein R1, R2, R3, R4, R5, R6, and R7 are as described herein. Methods are also provided for the synthesis of and use of the provided molecules in applications for diagnostic testing.

Abstract: Text can be encoded into DNA sequences. Each word from a document or other text sample can be encoded in a DNA sequence or DNA sequences and the DNA sequences can be stored for later retrieval. The DNA sequences can be stored digitally, or actual DNA molecules containing the sequences can be synthesized and stored. In one example, the encoding technique makes use of a polynomial function to transform words based on the Latin alphabet into k-mer DNA sequences of length k. Because the whole bits required for the DNA sequences are smaller than the actual strings of words, storing documents using DNA sequences may compress the documents relative to storing the same documents using other techniques. In at least one example, the mapping between words and DNA sequences is one-to-one and the collision ratio for the encoding is low.

Abstract: An amphoteric dissociation ion exchange separation medium, the surface of which is an amphoteric dissociation covalently-modified layer. When an environmental pH value is lower than the isoelectric point, pIm, of the covalently-modified layer, the type of net charges on the surface of the covalently-modified layer is positive and the separation medium has the properties of an anion exchanger; when the environmental pH value is higher than the pIm, the type of net charges on the covalently-modified layer surface is negative and the separation medium has the properties of a cation exchanger. The separation medium has the properties of an anion exchanger and a cation exchanger at both sides of the pIm, respectively. The pH of an eluent can be adjusted to allow the separation medium surface and the target substance to have the same type of net charges, so that the target substance can be released by electrostatic repulsion.

Abstract: The invention relates to the amplification of specific target nucleic acids. The invention provides methods, reagents, and kits for carrying out such amplification via the autoligation chain reaction (ACR).

Abstract: Provided herein is reagent mixture comprising multiplexed amplification reagents and flap assay reagents for detecting, in a single reaction, mutant copies of the KRAS gene that contain any of the 34A, 34C, 34T, 35A, 35C, 35T or 38A point mutations. Methods that employ the reagent mix and kits for performing the same are also provided.

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: The present disclosure provides methods of determining one or more tissues and/or cell-types contributing to cell-free DNA (“cfDNA”) in a biological sample of a subject. In some embodiments, the present disclosure provides a method of identifying a disease or disorder in a subject as a function of one or more determined more tissues and/or cell-types contributing to cfDNA in a biological sample from the subject.

Abstract: A method and system have been provided to perform high speed nucleic acid melting analysis while still obtaining accurate melting curve sufficient for genotyping. This rapid ability to interrogate DNA should be useful whenever time to result is important, such as in molecular point of care testing. Specifically, microfluidics enables genotyping by melting analysis at rates up to 50° C./s, requiring less than is to acquire an entire melting curve. High speed melting reduces the time for melting analysis, decreases errors, and improves genotype discrimination of small amplicons.

Abstract: A micro-plate reader for use with a portable electronic device having a camera includes an opto-mechanical attachment configured to attach/detach to the portable electronic device and includes an array of illumination sources. A slot in the opto-mechanical attachment is dimensioned to receive an optically transparent plate containing an array of wells. Optical fibers are located in the opto-mechanical attachment and transmit light from each well to a reduced size header having, wherein the fiber array in the header has a cross-sectional area that is ?10× the cross-sectional area of the wells in the plate. A lens located in the opto-mechanical attachment transmits light from the header fibers to the camera. Software executed on the portable electronic device or other computer is used to process the images to generate qualitative clinical determinations and/or quantitative index values for the separate wells.

Abstract: The invention is a novel method of making and using a library such as a sequencing library of single stranded circular nucleic acid templates via splint ligation. In particular, disclosed are methods of making circular target nucleic acid molecules and libraries of such molecules for downstream analysis such as nucleic acid sequencing. The method comprises the steps of adding universal sequences to nucleic acid molecules, rendering single-stranded these nucleic acid molecules with universal sequences on their ends by contacting with a probe complementary to at least a portion of the universal sequences, and allowing the hybridized probe to enable circularization and formation of single-stranded circular (sscDNA) molecules.

Abstract: The present invention relates to the field of pharmacogenomics and in particular to detecting the presence or absence of hypermethylated DNA. The detection of CpG methylation in marker DNA is useful for the diagnosis of cancers and the invention provides improved methods for this purpose. These improved methods allow in particular for a more sensitive detection of methylated marker DNA with high backgrounds of unmethylated marker DNA.

Abstract: High-fidelity, high-throughput nucleic acid sequencing enables healthcare practitioners and patients to gain insight into genetic variants and potential health risks. However, previous methods of nucleic acid sequencing often introduce sequencing errors (for example, mutations that arise during the preparation of a nucleic acid library, during amplification, or sequencing). Provided herein are methods and compositions for sequencing nucleic acids. Further provided are methods of identifying an error in a nucleic acid sequence.

Abstract: A method for identifying a nucleotide in a template nucleic acid by (a) providing a plurality of primer-template nucleic acid hybrids, wherein the primers have an extendable 3? end; (b) contacting the plurality with: (i) blocked nucleotides to produce a first subset of the primer-template nucleic acid hybrids that include a blocked nucleotide at the 3? end, and (ii) a ternary complex inhibitor to produce a second subset of the primer-template nucleic acid hybrids that include a ternary complex inhibitor; (c) forming ternary complexes that each include a polymerase, a primer-template nucleic acid hybrid of the first subset, and a cognate nucleotide; and (d) detecting the ternary complexes, thereby identifying a nucleotide in the template nucleic acid.

Abstract: The present invention relates to DNA microscopy methods to record the cellular co-localization and/or spatial distributions of arbitrary nucleic acid sequences, or other biomolecules tagged with nucleic sequences. The method involves sequence-components which may identify the targeted sequences-of-interest themselves and/or spatial beacons relative to which their distances are measured.

Abstract: The invention provides compositions and methods for the diagnosis, treatment, assessment, and characterization of hyperlipidemia-related diseases and disorders, including atherosclerosis, non-alcoholic fatty liver disease, obesity and diabetes mellitus in a subject in need thereof, based on the expression level of at least one miRNA that is associated with these diseases and disorders.

Abstract: The present disclosure provides compositions, methods and systems for quantifying target sequences and identifying target sequence variants.

Abstract: The invention relates to a method for the duplication of nucleic acids by means of a polymerase chain reaction, in the case of which a cycle consisting of the steps of denaturing, annealing and elongation is repeatedly performed. In one embodiment, in at least one passage of the cycle, the quotient of the duration of effect tA and the reaction volume Vr irradiated by the energy source is less than 1 seconds per microliter. In another embodiment, in at least one passage of the cycle, the ratio of the duration of effect (tA) and the duration of the PCR cycle (tc) is less than 20%. In certain embodiments, the yield (g) of nucleic acids at the end of at least one of the passages of the cycle is less than 80% of the nucleic acids present at the start of the passage.

Abstract: The presently disclosed subject matter provides for in vitro methods of inducing differentiation of human stem cells into neural crest, cranial placode or non-neuro ectoderm precursors, and cells generated by such methods. The presently disclosed subject matter also provides for uses of such cells for treating neurodegenerative and pituitary disorders.

Abstract: The present invention provides assays systems and methods for detection of chromosomal abnormalities and status of single loci associated with monogenic or polygenic traits in a sample containing nucleic acids from a maternal and a fetal source.

Abstract: Embodiments of the present invention relate to sequencing nucleic acids. In particular, embodiments of the methods and compositions provided herein relate to preparing nucleic acid templates and obtaining sequence data therefrom.

Abstract: Disclosed are methods for sequencing immune cell receptor repertoires from immune cell populations, the methods comprising isolating RNA from immune cells, generating cDNA from the RNA, ligating adapter sequences to the cDNA, and sequencing the cDNA. Also provided are kits containing primer mixtures for the sequencing of immune cell receptor repertoires.

Abstract: A method of designing a D-polypeptide that binds with an L-target protein can include: identifying a polypeptide target having L-chirality; determining hotspot amino acids of a polypeptide ligand having L-chirality that have binding interactions with the L-target protein; determining transformations of side chains of the hotspot amino acids that retain the binding interactions with the target; generating inversed hotspot amino acids with chirality opposite to the one of the target; identifying a polypeptide having inverse chirality from the target protein, on which a combination of inversed hotspot amino-acid can be grafted without significantly changing their interactions with the target. The designed ligands can be processed and converted to D-ligands that bind with the L-target protein.