Abstract: The invention relates to a method for synthesising long nucleic acids, including at least one cycle of elongating initial fragments of nucleic acids, including a) a phase comprising the enzymatic addition of nucleotides to said fragments, b) a phase comprising the purification of the fragments having a correct sequence, c) an optional phase of enzymatic amplification, each cycle being performed in a reaction medium which is compatible with enzymatic addition and amplification, such as an aqueous medium, the synthesis method also comprising, at the end of all the elongation cycles, a last step of final amplification. The invention also relates to the use of the method for the production of genes, or sequences of synthetic nucleic acids, DNA or RNA. The invention further relates to a kit for implementing said method.

Abstract: The present invention relates to a genetically-engineered Mycobacterium strain and a use thereof in the preparation of steroidal compounds. The genetically-engineered Mycobacterium strain is a Mycobacteria which lacks of acyl-CoA dehydrogenase genes fadE31, fadE32 and fadE33, wherein acyl-CoA dehydrogenase genes fadE31, fadE32 and fadE33 respectively encode proteins as follows: having amino acid sequences according to SEQ ID NOs 4, 6 and 8; derived by substituting, deleting or inserting one or more amino acids in the amino acid sequence defined by preceding protein and having the same function as that of the preceding protein. The present invention constructs a genetically-engineered Mycobacterium strain and applies it in preparing steroidal compounds, thereby enriching the types of valuable intermediates, improving the production efficiency and product quality of steroid drugs, reducing energy consumption in the steroid drugs production, simplifying production steps, and reducing production costs.

Abstract: The present invention provides a method for determining whether or not a test sample contains a phytopathogenic fungielectively from two kinds of fungi of a phytopathogenic fungus and a non-phytopathogenic fungus. The method according to the present invention comprises: (a) putting the test sample on a front surface of a substrate comprising a through hole; wherein the substrate comprises a cellulose film on a back surface thereof; the cellulose film has no through hole; the cellulose film has a thickness of more than 2 micrometers and not more than 3.7 micrometers; and the through hole has a cross-sectional area of not less than 7.065 square micrometers and not more than 19.625 square micrometers; (b) leaving the test sample at rest; (c) observing a back surface of the cellulose film; and (d) determining that the test sample contains the phytopathogenic fungus, if a fungus which has penetrated the cellulose film is found on the back surface of the cellulose film.

Abstract: The present disclosure relates to a method for manufacturing a three-dimensional cell culture support having a double crosslink, and a casting tray for manufacturing the three-dimensional cell culture support, wherein the method for manufacturing the three-dimensional cell culture support having the double crosslink includes: producing a cell mixed hydrogel; manufacturing a casting gel mold in a three-dimensional shape; and manufacturing a structure gelated in a three-dimensional shape, and the casting tray for manufacturing the three-dimensional cell culture support includes: a tray part including a groove accommodating a gel solution; a mold part covering the tray part; and a mold protrusion provided on the mold part and inserted into the groove when the mold part covers the tray part.

Abstract: Disclosed are methods and compositions which may be used in human cytochrome P450 (CYP450) enzyme phenotyping. The methods and compositions typically utilize a mélange of substrates for different CYP450 enzymes which may be administered orally to a patient. Subsequently, the metabolites of the substrates may be detected in the patient's saliva as well as any non-metabolized substrates to calculate a metabolic ratio for any given CYP450 enzyme in order to generate a phenotypic CYP450 enzyme profile for the patient.

Abstract: The invention relates to biosensors. More particularly, this invention relates to an electrochemical biosensor and to electrochemically active enzymes or variants thereof that are suitable for detection of one or more target molecules in a sample.

Abstract: Non-caloric artificial sweeteners (NAS) are used as a substitute for natural sugars by providing the sweet taste. This invention measures the effects of artificial sweeteners on the enzyme kinetics of biological systems. The claimed invention is directed to a method of measuring the effect of an artificial sweetener on enzyme-catalyzed hydrolysis of a sugar comprising establishing a first sugar enzymatic conversion rate, then determining a second sugar enzymatic conversion rate, and lastly comparing the first sugar enzymatic rate and the second sugar enzymatic conversion rate. The sugar enzymatic conversion rates can be measured by either nuclear magnetic resonance spectroscopy or a glucometer. The methodology presented may be applied to the elucidation of kinetic parameters for invertase catalyzed conversion of sucrose to glucose and fructose.

Abstract: A method for determining the concentration of an analyte in a sample, and to a test strip and a kit for carrying out said method, and a test system having said test strip and a detector. The use of the method to determine the concentration of an analyte in a sample.

Abstract: The present invention relates to a method for assessing the status of nucleic acid degradation and/or integrity of one or more nucleic acids in a sample, comprising the steps amplifying at least two overlapping regions within at least one locus (e.g. by heminested or nested PCR), and detecting the amount of the at least two amplification products through the use of at least two probes, wherein one probe binds to the region of overlap and the at least one other probe binds to a non-overlapping region. The invention further relates to a method of designing primers and/or probes for amplifying at least two overlapping regions within at least one locus, wherein the locus that is amplified is a single copy locus (SCL) or multicopy locus (MLC). The invention also relates to a primer and a primer pair for amplifying at least two overlapping regions from one nucleic acid template which is a multicopy locus present in 21 loci in the human genome. These primers and probes may be in a kit.

Abstract: The present invention and embodiments thereof relates to compositions and methods for storage, stabilization and preservation of biological samples and/or nucleic acids on a solid matrix. Methods for extracting, collecting, and recovering the biological samples and/or nucleic acids from the solid matrix are also described.

Abstract: Provided is a method for obtaining base sequence information of a single cell derived from a vertebrate, by which PCR amplification of an objective region can be performed uniformly and accurately.

Abstract: Provided are a method for discriminating an origin of human genomic DNA of 100 pg or less by uniformly amplifying human genomic DNA of 100 pg or less, a method for identifying an individual, and a method for analyzing a level of engraftment of hematopoietic stem cells.

Abstract: The present disclose provides methods and systems for amplifying and quantifying nucleic acids and for detecting the presence or absence of a target in a sample. The methods and systems provided herein may utilize a device comprising a plurality of partitions separated from an external environment by a gas-permeable barrier. Certain methods disclosed herein involve subjecting nucleic acid molecules in the plurality of partitions to conditions sufficient to conduct nucleic acid amplification reactions. The nucleic acid molecules may be subjected to controlled heating in the plurality of partitions to generate data indicative of a melting point(s) of the nucleic acid molecules.

Abstract: A method and a kit for detecting mycobacterium are provided. The method includes steps of: providing a sample; providing a pair of primers, which is selected from a group consisting of a sequence having about 45% to 100% similar to SEQ ID NO. 1, a sequence having about 60% to 100% similar to SEQ ID NO. 2, a sequence complementary thereof; performing a polymerase chain reaction by using the set of primers and the sample to obtain a product; and analyzing the product to detect the presence of the mycobacterium.

Abstract: The present invention relates to markers being capable of predicting or diagnosing the risk of metabolic syndrome or metabolic syndrome-related diseases and their use. More particularly, the present invention relates to a composition, a kit and a method for predicting or diagnosing the risk of metabolic syndrome or metabolic syndrome-related diseases by detecting Neisseria spp., Granulicatella spp., and/or Peptococcus spp. in a test sample.

Abstract: Systems and methods to assess the health of various microbiomes and to identity species therein are disclosed. Described assessments and identifications can inform treatment decisions if a microbiome is determined to have a less than optimal balance of bacterial species within it; the presence of one or more negative species; and/or the absence of one or more positive species.

Abstract: Systems and method for analyzing molecules in a sample. The system includes an imager, a flow cell, a magnet, and a processor. The flow cell includes a functionalized surface comprising a capture probe configured to bind molecules comprising a first RNA transcript. The magnet is positioned opposite the functionalized surface. The magnet is configured to direct the molecules comprising the first RNA transcript to the functionalized surface to bind to the capture probe. The light source configured to direct a light beam at the bound molecules comprising the first RNA transcript. The imager is configured to capture light from the bound molecules comprising the first RNA transcript. A processor configured to determine a quantity of the molecules in the sample comprising the first RNA transcript. The process is further configured to determine an expression level of the first RNA transcript in the sample based on the quantity of the molecules.

Abstract: The present invention provides methods and compositions involving genetic markers and their association with oral mucositis occurring in patients who are treated with cancer therapy and/or conditioning therapy for hematopoietic stein cell transplantation.

Abstract: Provided is a method of analyzing microhaplotypes by using a next generation sequencing (NGS). The method of analyzing microhaplotypes by using the NGS includes: (a) performing a multiplex PCR for simultaneously amplifying the microhaplotypes; (b) performing an indexing PCR by using a product of the multiplex PCR; and (c) performing the NGS by using a product of the indexing PCR.

Abstract: Among other things, the present disclosure pertains to methods and compositions related to phasing of allelic variants of genetic loci. Phasing of allelic variants of genetic loci on an individual patient's chromosomes is highly valuable for many purposes, including patient stratification for allele-specific therapeutics.

Abstract: Localized detection of RNA in a tissue sample that includes cells is accomplished on an array. The array include a number of features on a substrate. Each feature includes a different capture probe immobilized such that the capture probe has a free 3? end. Each feature occupies a distinct position on the array and has an area of less than about 1 mm2. Each capture probe is a nucleic acid molecule, which includes a positional domain including a nucleotide sequence unique to a particular feature, and a capture domain including a nucleotide sequence complementary to the RNA to be detected. The capture domain can be at a position 3? of the positional domain.

Abstract: The present invention provides a novel method for labelling nucleic acid probes. The method uses a ligase catalysed reaction to connect a nucleic acid probe with pre-prepared nucleic acid label carrier molecules under the presence of a stabilizing complementary splint oligonucleotide. The method allows for an easy, cheap and fast labelling of multiple probes with multiple different labels. In this way, the costs and effort for the generation of single molecule Fluorescent In Situ Hybridization (smFISH) assays was significantly reduced. The invention further provides methods for the generation of FISH libraries and labelling kits comprising the novel tools of the invention.

Abstract: The present invention generally relates to systems and methods for imaging or determining nucleic acids or other desired targets, for instance, within cells or tissues. In one aspect, a sample is exposed to a plurality of nucleic acid probes that are determined within the sample. In some cases, however, background fluorescence or off-target binding may make it more difficult to determine properly bound nucleic acid probes. Accordingly, other components of the samples that may be contributing to the background, such as proteins, lipids, and/or other non-targets, may be “cleared” from the sample to improve determination. However, in certain embodiments, nucleic acids or other desired targets may be prevented from also being cleared, e.g., using polymers or gels within the sample. Other aspects are generally directed to compositions or kits involving such systems, methods of using such systems, or the like.

Abstract: Embodiments of the disclosure encompass methods of amplifying nucleic acid from one or more cells. In particular embodiments, the nucleic acid is amplified as amplicons in a linear manner. Specific embodiments include the removal or effective destruction of nonlinearly produced amplicons.

Abstract: Novel methods of generating a localized population of immobilized clonal amplicons on a support are provided.

Abstract: The invention generally relates to sequencing library preparation methods. In certain embodiments, two or more template nucleic acids are joined together by a linking molecule, such as a PEG derivative. Identical copies of a nucleic acid fragment or both strands of a duplex fragment may be linked together. The linked nucleic acids are amplified, creating linked amplicons. Emulsion PCR with linked primers creates linked template nucleic acids for seeding sequencing clusters and errors can be readily identified by their presence on only one of the linked fragments.

Abstract: A method of preparing reagents includes inserting a cartridge into an instrument. The cartridge includes a plurality of reagent enclosures disposed in a cavity of the cartridge and exposing a port to an exterior of the cartridge. Each reagent enclosure includes a reagent container including a reagent and an internal cavity defining a compressible volume, an opening defined through the reagent container to the internal cavity. The method further includes connecting a plurality of fluid ports to the openings of the plurality of reagent enclosures; applying a solution through the fluid ports to at least partially fill the plurality of reagent enclosures; and cycling a pressure of the cavity, whereby for each of the reagent enclosures, during increasing pressure, the solution enters the internal cavity of the reagent container, combines with the reagent, and compresses the compressible volume, and during decreasing pressure, the compressible volume decreases and the reagent is ejected through the opening.

Abstract: The present invention relates to a method of using nanopores to obtain sequence information of sample DNAs in ss test DNAs. The method comprises using speed bumps to stall the ss test DNAs in the nanopores at random positions of the ss test DNAs to obtain sequence information of each and every nucleotides of the sample DNAs, and to construct the whole sequences of the sample DNAs. The present invention also relates to identification and/or isolation of test DNAs having desired sequence(s) using nanopore detectors facilitated by speed bump.

Abstract: A stack of fluidics layers of a microfluidic cartridge for sequencing nucleic acid molecules includes a sequencing chamber layer having a sequencing chamber area configured for carrying out clustering and sequencing reactions, and a sequencing chamber bottom layer disposed under the sequencing chamber layer. The sequencing chamber bottom layer has an opening configured to hold an image sensor with the image sensor having an active area disposed under the sequencing chamber area. The sequencing chamber area spans substantially all of the active area of the image sensor. The stack of fluidics layers includes a flexible printed circuit board (PCB) layer under the sequencing chamber bottom layer, and a fluidics channels layer disposed under the flexible PCB layer. The fluidics channels layer includes fluidics channels that are configured to deliver reactants to the sequencing chamber area. The fluidics channels do not substantially overlap with the active area of the image sensor.

Abstract: The present disclosure provides methods for determining the ploidy status of a chromosome in a gestating fetus from genotypic data measured from a mixed sample of DNA comprising DNA from both the mother of the fetus and from the fetus, and optionally from genotypic data from the mother and father. The ploidy state is determined by using a joint distribution model to create a plurality of expected allele distributions for different possible fetal ploidy states given the parental genotypic data, and comparing the expected allelic distributions to the pattern of measured allelic distributions measured in the mixed sample, and choosing the ploidy state whose expected allelic distribution pattern most closely matches the observed allelic distribution pattern. The mixed sample of DNA may be preferentially enriched at a plurality of polymorphic loci in a way that minimizes the allelic bias, for example using massively multiplexed targeted PCR.

Abstract: The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing. Such polynucleotide processing may be useful for a variety of applications, including polynucleotide sequencing.

Abstract: The present disclosure provides methods and systems for detecting multiple different nucleotides in a sample. In particular, the disclosure provides for detection of multiple different nucleotides in a sample utilizing fewer detection moieties than the number of nucleotides being detected and/or fewer imaging events than the number of nucleotides being detected.

Abstract: Disclosed herein is a system and method for increasing the fidelity of measured genetic data, for making allele calls, and for determining the state of aneuploidy, in one or a small set of cells, or from fragmentary DNA, where a limited quantity of genetic data is available. Poorly or incorrectly measured base pairs, missing alleles and missing regions are reconstructed using expected similarities between the target genome and the genome of genetically related individuals. In accordance with one embodiment, incomplete genetic data from an embryonic cell are reconstructed at a plurality of loci using the more complete genetic data from a larger sample of diploid cells from one or both parents, with or without haploid genetic data from one or both parents. In another embodiment, the chromosome copy number can be determined from the measured genetic data, with or without genetic information from one or both parents.

Abstract: The present invention relates to a method for discriminating an operationally tolerant (TOL) subject from a non-operationally tolerant (STA) subject, comprising the following steps: i) establishing a composite score of tolerance (cSoT) with the expression levels of six genes in a biological sample obtained from said subject and two clinical parameters; wherein said six genes are ID3, AKR1C3, CD40, CTLA4, TCL1A and MZB1, and wherein said cSoT is established by the following formula: cSoT = ? i n ? = ? i × Exprs + ? test ? ? time × age test ? ? time + ? trans ? ? time × age trans ? ? time + intercept - scaling ? ? coefficient ii) comparing this cSoT with a predetermined reference value; and iii) concluding that the subject is TOL when the cSoT is higher than the predetermined reference value or concluding that the subject is STA when the cSoT is lower than the predetermined reference value.

Abstract: The present disclosure relates to the identification of single nucleotide polymorphisms (SNPs) in the Gamma genomic block in the central region of the major histocompatibility complex (MHC) that can be used for matching transplant donors and recipients and determining disease susceptibility.

Abstract: Embodiments of the invention relate generally to methods of identifying subjects likely to develop diabetes-associated damage to the nephron, or subjects in the early stages of diabetic nephropathy. In particular, several embodiments relate to quantification of diabetic nephropathy-associated RNA isolated from vesicles from patient urine samples is performed to compare a subject to a population having normal nephron function and/or to track progression of diabetic nephropathy in said subject over time.

Abstract: This application discloses methods for cDNA synthesis with improved reverse transcription, template switching and preamplification to increase both yield and average length of cDNA libraries generated from individual cells. The new methods include exchanging a single nucleoside residue for a locked nucleic acid (LNA) at the TSO 3? end, using a methyl group donor, and/or a MgCl2 concentration higher than conventionally used. Single-cell transcriptome analyses incorporating these differences have full-length coverage, improved sensitivity and accuracy, have less bias and are more amendable to cost-effective automation. The invention also provides cDNA molecules comprising a locked nucleic acid at the 3?-end, compositions and cDNA libraries comprising these cDNA molecules, and methods for single-cell transcriptome profiling.

Abstract: Disclosed are methods, kits, and devices for diagnosing and treating psychiatric disorders and the symptoms thereof. The methods, kits, and devices relate to identifying genetic markers that may be utilized to diagnose and/or prognose a subject and treat the diagnosed and/or prognosed subject by administering a drug the subject based on the genetic marker having been identified. Genetic markers identified in the methods may include a polymorphism in a gene encoding a protein associated with synaptogenic adhesion, scaffolding, neuron-specific splicing regulation, potassium channels which form leak conductances that regulate neuronal excitability, synaptic spine turnover and stability of synaptic contacts, and/or vesicle trafficking and exocytosis in presynaptic neurons and neuromuscular junctions.

Abstract: Methods and compositions are provided for detecting a predisposition for cardiovascular disease in an individual.

Abstract: A method includes obtaining a biological sample from an individual, extracting DNA from cells in the sample, measuring the degree of methylation of the extracted DNA at one or more specified loci, and identifying if at least one locus exhibits an alteration in methylation or a degree of methylation that is outside of a normal limit. The method may further include determining whether an individual was exposed to abuse and/or trauma. In some cases, the method further includes treating the individual with a therapy effective to reverse the differential methylation of the DNA.

Abstract: A system and method for determining the genetic data for one or a small set of cells, or from fragmentary DNA, where a limited quantity of genetic data is available, are disclosed. Genetic data for the target individual is acquired and amplified using known methods, and poorly measured base pairs, missing alleles and missing regions are reconstructed using expected similarities between the target genome and the genome of genetically related subjects. In accordance with one embodiment of the invention, incomplete genetic data is acquired from embryonic cells, fetal cells, or cell-free fetal DNA isolated from the mother's blood, and the incomplete genetic data is reconstructed using the more complete genetic data from a larger sample diploid cells from one or both parents, with or without genetic data from haploid cells from one or both parents, and/or genetic data taken from other related individuals.

Abstract: Provided are methods and assays for cancer cell classification, cancer prognosis and treatment based on the isolation of circulating tumor cells and the characterization of their nuclear organization and telomere signatures.

Abstract: Methods for classifying and for evaluating the prognosis of a subject having breast cancer are provided. The methods include prediction of breast cancer subtype using a supervised algorithm trained to stratify subjects on the basis of breast cancer intrinsic subtype. The prediction model is based on the gene expression profile of the intrinsic genes listed in Table 1. This prediction model can be used to accurately predict the intrinsic subtype of a subject diagnosed with or suspected of having breast cancer. Further provided are compositions and methods for predicting outcome or response to therapy of a subject diagnosed with or suspected of having breast cancer. These methods are useful for guiding or determining treatment options for a subject afflicted with breast cancer.

Abstract: A frequency of somatic mutations in a biological sample (e.g., plasma or serum) of a subject undergoing screening or monitoring for cancer, can be compared with that in the constitutional DNA of the same subject. A parameter can derived from these frequencies and used to determine a classification of a level of cancer. False positives can be filtered out by requiring any variant locus to have at least a specified number of variant sequence reads (tags), thereby providing a more accurate parameter. The relative frequencies for different variant loci can be analyzed to determine a level of heterogeneity of tumors in a patient.

Abstract: The present disclosure relates to a use of H2AFZ as a hepatocellular carcinoma (HCC) biomarker and more particularly, to a marker for diagnosing hepatocellular carcinoma consisting of a H2AFZ gene or an expression protein H2A.Z.1 thereof, a composition for diagnosing or estimating prognosis of HCC, a method for diagnosing or estimating prognosis of HCC, a method of detecting a biomarker for diagnosing or estimating prognosis of HCC, a screening method of an HCC therapeutic agent, and a pharmaceutical composition for preventing or treating HCC.

Abstract: The present disclosure is directed to a streamlined sample preparation process, VERA (Viral Enrichment by Reducing Artifacts), to tilt total genomic material in favor of DNA/RNA viral genomes. This reduction of host genomic artifacts can be completed in <8 hours from sample acquisition. Using a rapid library preparation protocol (˜1.5 minutes) and real-time nanopore sequencing, potential viral contamination, for example RNA viral contamination, can be identified in less than one workday from sample acquisition.

Abstract: The disclosure describes methods, kits, apparatus and oligonucleotides for the detection of Zika virus. In one aspect, the disclosure describes a method of determining whether a sample comprises ZIKA virus (ZIKV) nucleic acid comprising performing an amplification reaction with the sample in the presence of a second oligonucleotide set, a first oligonucleotide set, a third oligonucleotide set or a combination of two or more of the sets, wherein: the first oligonucleotide set has: an oligonucleotide ON A with a consecutive stretch of at least 18 nucleotides of the sequence AARTACACATACCARAACAAAGTGGT (FP_ZIKA_San0-F) (SEQ ID NO:1) or a consecutive stretch of at least 18 nucleotides of the sequence TACACATACCARAACAAAGTGGT (FP_ZIKV_San0.

Abstract: The present invention discloses a novel endophytic fungi, Ovatospora brasiliensis MTCC 25236 for the bioconversion of curcuminoids to Calebin-A and a method for its isolation from the rhizomes of Curcuma sp. The invention also discloses a method for the bioconversion of curcuminoids to Calebin-A using an endophytic fungi Ovatospora brasiliensis MTCC 25236 and bacterial species, Acinetobacter johnsonii and Pseudomonas putida.

Abstract: A method for forming a vehicle component is provided. The method may include heating a blank of 36MnB5 in a furnace to an austenitization temperature, stamping the blank with a die assembly to form a vehicle component and change a microstructure of the blank from austenite to martensite, and responsive to a temperature of the vehicle component being at or below 130° C., removing the vehicle component from the die assembly such that the vehicle component has a yield strength equal to or greater than 1400 MPa. The blank may be retained within the die assembly for a quench time of between five and eleven seconds following the stamping. The method may further include thermo-stabilizing a surface of the die assembly to a predetermined temperature. The stamping may further include applying a pressure of approximately 10 N/mm2 to the blank.

Abstract: A steel product includes the following chemical composition in wt. %: C: 0.01 to <0.3, Mn: 4 to <10, Al: 0.003 to 2.9, Mo: 0.01 to 0.8, Si: 0.02 to 0.8, Ni: 0.005 to 3, P: <0.04, S: <0.02, N: <0.02, with the remainder being iron including unavoidable steel-associated elements, wherein an alloy composition satisfies the equation 6<1.5 Mn+Ni<8; or the equation 0.11<C+Al<3, or an alloy composition contains, in addition to Ni, at least one or more of the elements, in wt. %, B: 0.0005 to 0.014; V: 0.006 to 0.1; Nb: 0.003 to 0.1; Co: 0.003 to 3; W: 0.03 to 2 or Zr: 0.03 to 1. The steel product has a microstructure of 2 to 90 vol. % austenite, less than 40 vol. % ferrite and/or bainite, with the remainder being martensite.