Abstract: Systems, networks, methods, compositions and recombinant hosts for biosynthesizing hydrocarbons from a feedstock, such as gas, are provided.

Abstract: The present invention is directed to a polymerase activity assay that produces a strong optical signal when a primer-template complex is extended to full-length product. The assay uses Cy3 as the molecular beacon and Iowa Black® RQ as the quencher. The signal-to-noise-ratio (STNR) of this donor-quencher pairing is ˜200-fold over background, which is considerably better than other donor-quencher pairs (STNRs ˜10-20-fold). The STNR allows for solution-based monitoring of polymerase activity. Because the sensor functions via Watson-Crick base pairing, the polymerase activity assay may also be used to evolve polymerases to accept xeno nucleic acids as substrates.

Abstract: Deep mutational scanning is a foundational tool for addressing functional consequences of large numbers of mutants, yet a more efficient and accessible method for construction of user-defined mutagenesis libraries is needed. Provided herein are nicking saturation mutagenesis, a single-day, single-pot saturation mutagenesis method using routinely prepped plasmid dsDNA as input substrate. Reproducibility and convenience of the method are demonstrated through validation by an external research laboratory.

Abstract: The present technology provides for an apparatus for detecting polynucleotides in samples, particularly from biological samples. The technology more particularly relates to microfluidic systems that carry out PCR on nucleotides of interest within microfluidic channels, and detect those nucleotides. The apparatus includes a microfluidic cartridge that is configured to accept a plurality of samples, and which can carry out PCR on each sample individually, or a group of, or all of the plurality of samples simultaneously.

Abstract: The invention relates to a new method of characterising a target ribonucleic acid (RNA) involving forming a complementary polynucleotide. The method uses a transmembrane pore.

Abstract: An imaging agent comprises (a) at least one target recognition moiety; (b) at least one observable moiety non-transiently bound to the target recognition moiety, and (c) at least one docking moiety bound to the target recognition moiety, wherein the docking moiety is capable of transiently binding at least one observable moiety. In some embodiments, the at least one target recognition moiety is an antibody or antigen binding fragment thereof.

Abstract: Methods and apparatus relate to the synthesis of high fidelity polynucleotides and to the reduction of sequence errors generated during synthesis of nucleic acids on a solid support. Specifically, design of support-bound template oligonucleotides is disclosed. Assembly methods include cycles of annealing, stringent wash and extension of polynucleotides comprising a sequence region complementary to immobilized template oligonucleotides. The error free synthetic nucleic acids generated therefrom can be used for a variety of applications, including synthesis of biofuels and value-added pharmaceutical products.

Abstract: A method of diagnosing or predicting a bladder cancer, or a risk of a developing a bladder cancer in a subject is provided, which method includes the detection of specific mutations in the FGFR3 gene in a first biological sample; and the measure of the degree of methylation of target genes in a biological sample obtained from the subject in a second biological sample.

Abstract: A method and kit for the capture and purification of specific nucleic acids from a sample with affinity capture probes on a solid support and for the replication of said nucleic acids with a strand displacing polymerase, whereby a second primer complementary to a sequence in each of the target nucleic acids distinct from that bound by capture probes is also bound to the nucleic acid targets, and extension of one of the primers on each target effects the separation of the copied nucleic acid strands from the solid support. Incorporation of universal nucleic acid sequences during their replication enables the simultaneous and highly specific amplification of multiple nucleic acid target sequences with minimal production of artifacts.

Abstract: Methods for the rapid detection of the presence or absence of Mycobacterium tuberculosis (MTB) and other members of the MTB-complex in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers, probes targeting the esxJ gene, along with kits are provided that are designed for the detection of MTB.

Abstract: The present invention provides a system for receiving biological sequence information and activating the synthesis of a biological entity. The system has a receiving unit for receiving a signal encoding biological sequence information transmitted from a transmitting unit. The transmitting unit can be present at a remote location from the receiving unit. The system also has an assembly unit connected to the receiving unit, and the assembly unit assembles the biological entity according to the biological sequence information. Thus, according to the present invention biological sequence information can be digitally transmitted to a remote location and the information converted into a biological entity, for example a protein useful as a vaccine, immediately upon being received by the receiving unit and without further human intervention after preparing the system for receipt of the information.

Abstract: Transposomes and oligonucleotide replacement methods to make DNA libraries that have distinct 5? and 3? tags, and to make directional libraries that are enriched for a desired strand.

Abstract: Sharing data is disclosed. In some cases, sharing data includes receiving a request to share data from a first account to a second account, receiving an indication of a plurality of first account profiles associated with the first account to share with the second account, and establishing sharing from the plurality of first account profiles to the second account, wherein sharing comprises the second account having read access to a subset of nonpublic data associated with the plurality of first account profiles.

Abstract: Embodiments provided herein relate to methods and compositions for preparing nucleic acid libraries. Some embodiments include preparing libraries from nucleic acids obtained from degraded samples, such as ancient samples and fixed samples.

Abstract: This invention relates to mutant DNA polymerases having an enhanced template discrimination activity compared with the corresponding unmodified DNA polymerase counterparts, wherein the amino acid sequence of the mutant DNA polymerase includes at least one substitution at residue positions structurally and functionally homologous or orthologous positions 783 or 784 of an unmodified Taq DNA polymerase.

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 blocking the 3? ends of pooled indexed polynucleotides from multiple samples prior to amplification and sequencing.

Abstract: The present disclosure provides methods and systems for amplifying and analyzing nucleic acid samples. The present disclosure provides methods for preparing cDNA and/or DNA molecules and cDNA and/or DNA libraries using modified reverse transcriptases.

Abstract: The present disclosure provides novel primers and method for the detection of specific nucleic acid sequences. The primers and methods provided herein are useful in a wide variety of molecular biology applications and are particularly useful in allele-specific PCR.

Abstract: Methods, systems, and reagents for real-time single molecule sequencing of nucleic acids, particularly long DNA molecules, is described. Preferably, such methods and systems combine FRET (Förster Resonance Energy Transfer)-based proximity sensing of labeled nucleotides in or near a DNA polymerase's active site with FRET-based monitoring of the conformational changes in the polymerase that occur during nucleotide incorporation.

Abstract: Disclosed is a nucleic acid amplification method. The method comprises: performing a first nucleic acid amplification using a target nucleic acid contained in a sample as a template; and performing a second nucleic acid amplification using the amplification product produced in the first nucleic acid amplification step as a template. In the first nucleic acid amplification, a DNA polymerase that selectively amplifies a nucleic acid not comprising a uracil base is used to produce an amplification product that does not comprise a uracil base. In the second nucleic acid amplification, (i) dUTP and/or a primer comprising a uracil base, and (ii) a DNA polymerase capable of amplifying a nucleic acid comprising a uracil base are used to produce an amplification product that comprises a uracil base.

Abstract: Described is a method for the production isoamyl alcohol (3-methylbutan-1-ol) comprising the enzymatic conversion of 3-methylbutyryl-CoA (isovaleryl-CoA) into isoamyl alcohol comprising: (a) two enzymatic steps comprising (i) first the enzymatic conversion of 3-methylbutyryl-CoA into 3-methylbutyraldehyde (3-methylbutanal or isovaleraldehyde); and (ii) then enzymatically converting the thus obtained 3-methylbutyraldehyde into said isoamyl alcohol; or (b) a single enzymatic reaction in which 3-methylbutyryl-CoA is directly converted into isoamyl alcohol by making use of an alcohol-forming short chain acyl-CoA dehydrogenase/fatty acyl-CoA reductase or an alcohol-forming fatty acyl-CoA reductase (long-chain acyl-CoA:NADPH reductase) (EC 1.2.1.84). Further, described is the above method wherein the 3-methylbutyryl-CoA can be provided by the enzymatic conversion of 3-methylcrotonyl-CoA into said 3-methylbutyryl-CoA.

Abstract: The present invention relates to a method and kit for analyte detection. More precisely the invention relates to a method of detecting an analyte, comprising storing short single stranded nucleic acids (10-100 nt), such as aptamers and micro RNA, on a solid support at ambient temperature; and subsequent amplification of said nucleic acids for detection of analyte(s).

Abstract: The present disclosure is directed to a kit for the detection of an analyte of interest in a sample comprising, in one or more vials, containers, or compartments, a. a surface comprising (i) a capture reagent for the analyte, said capture reagent bound to the surface, and (ii) an anchoring reagent bound to an anchoring oligonucleotide sequence, said anchoring reagent bound to the surface, b. a first detection reagent for the analyte that is linked to a first nucleic acid probe, wherein the first nucleic acid probe comprises an extended sequence that is complementary to the anchoring oligonucleotide sequence and c. a second detection reagent for the analyte that is linked to a second nucleic acid probe.

Abstract: A portable pathogen analysis system (PPAS) designed to detect microbial pathogens at the point-of-sample collection. The system comprises a concentration tube used for the concentration of microbes in large volumes of water samples using super absorbent polymer (SAP) beads and a hand-powered centrifuge; and a processing component, which functions as a portable lab-on-a-disc droplet digital nucleic acid amplification system, which integrates DNA extraction, nucleic acid amplification, and post-amplicon analysis in a single unit. The present invention provides a fast, cost-effective, and user-friendly solution for microbial water quality analysis in low-resource settings.

Abstract: Examples include polymerase chain reaction (PCR) devices. Example PCR devices comprise a fluid input, ejection nozzles, and a set of microfluidic channels that fluidly connect the fluid input and the ejection nozzles. Each microfluidic channel comprises a reaction chamber, and examples further comprise at least one heating element, where the at least one heating element is positioned in the reaction chamber of each microfluidic channel. The at least one heating element is to heat fluid in the reaction chamber of each fluid channel. The device may eject fluid via the ejection nozzles.

Abstract: The present disclosure provides a system and method for the detection of rare mutations and copy number variations in cell free polynucleotides. Generally, the systems and methods comprise sample preparation, or the extraction and isolation of cell free polynucleotide sequences from a bodily fluid; subsequent sequencing of cell free polynucleotides by techniques known in the art; and application of bioinformatics tools to detect rare mutations and copy number variations as compared to a reference. The systems and methods also may contain a database or collection of different rare mutations or copy number variation profiles of different diseases, to be used as additional references in aiding detection of rare mutations, copy number variation profiling or general genetic profiling of a disease.

Abstract: The invention relates to a sensor arrangement for a motor vehicle, with a pane, a sensor housing, and an optical sensor, wherein the sensor housing is arranged on one side of the pane such that between the sensor housing and the pane, a gap is formed, in which the optical sensor is arranged. To allow for a deicing of the pane in an area of the sensor housing in a particularly simple and thus cost-effective manner, it is provided that between the sensor housing and the side of the pane, a heat conducting element is arranged to conduct heat from outside the sensor housing in between the sensor housing and the pane.

Abstract: The present disclosure relates to a rapid diagnostic test system that includes the prospective collection of whole blood, preservation of circulating nucleic acids at ambient temperature, and the reproducible detection of nucleic acids including DNA and mRNA (including fusion transcripts and differentially expressed transcripts) by different genomic methodologies.

Abstract: The invention provides compositions and methods for modifying the 3?-terminal ends of nucleic acids using DNA polymerase ? terminal transferase activity.

Abstract: In one aspect, the invention provides a method of screening a human subject to determine if said subject has a genetic predisposition to develop, or is suffering from Facioscapulohumeral Dystrophy (FSHD), said method comprising: (a) providing a biological sample comprising genomic DNA from the subject; and (b) analyzing the portion of the genomic DNA in the sample corresponding to the distal D4Z4-pLAM region on chromosome 4 and determining the presence or absence of a polymorphism resulting in a functional polyadenylation sequence operationally linked to exon 3 of the DUX4 gene, wherein a determination of the absence of a functional polyadenylation sequence operationally linked to exon 3 of the DUX4 gene indicates that the subject does not have a genetic predisposition to develop, and is not suffering from FSHD, and/or wherein a determination of the presence of a functional polyadenylation sequence operationally linked to exon 3 of the DUX4 gene indicates that the subject has a genetic predisposition to devel

Abstract: A hermetically sealed multi-axis articulated arm apparatus for use within a sealable isolator chamber comprises a rotational shaft that passes through an opening in the chamber, a sealing member disposed within the chamber for sealing the shaft to an inner surface of the chamber, a plurality of interconnected hermetically sealed arm segments, operably attached to the linear motion shaft; an end effector operably attached to a terminal arm segment among the plurality of arm segments, and at least one fully enclosed drive system for driving and controlling the shaft and the plurality of arm segments. The linear motion shaft may have a sealing member in the form of a bellows. The materials for the parts of the apparatus exposed to the atmosphere of the chamber are compatible with an aseptic and cleanable environment and the surfaces of the arm segments are shaped to avoid pooling of contaminants.

Abstract: The invention describes composition and methods of use for novel hairpin blocked-cleavable primers. In one embodiment unblocking occurs through action of RNase H2. The method improves the specificity of PCR and reduces primer dimer events, enabling higher level multiplex reactions. Additionally, the invention protects RNA-containing primers from attack by single-strand RNases.

Abstract: The present invention regards a variety of methods and compositions for whole genome amplification and whole transcriptome amplification. In a particular aspect of the present invention, there is a method of amplifying a genome comprising a library generation step followed by a library amplification step. In specific embodiments, the library generating step utilizes specific primer mixtures and a DNA polymerase, wherein the specific primer mixtures are designed to eliminate ability to self-hybridize and/or hybridize to other primers within a mixture but efficiently and frequently prime nucleic acid templates.

Abstract: Sharing data is disclosed. In some cases, sharing data includes receiving a request to share data from a first account to a second account, receiving an indication of a plurality of first account profiles associated with the first account to share with the second account, and establishing sharing from the plurality of first account profiles to the second account, wherein sharing comprises the second account having read access to a subset of nonpublic data associated with the plurality of first account profiles.

Abstract: This invention relates to compositions and methods for the differential detection of multiple viruses using a one-step assay. The viruses to be detected include Zika, West Nile, dengue (genotype 1-4) and chikungunya viruses. In particular, the invention relates to a method of and assay for differential detection of the viruses using specific primers and probes designed to detect and differentiate between the viruses.

Abstract: A method of detecting a predisposition to, or the incidence of, cancer in a sample comprises detecting an epigenetic change in at least one gene selected from an NDRG4/NDRG2 subfamily gene, GATA4, OSMR, GATA5, SFRP1, ADAM23, JPH3, SFRP2, APC, MGMT, TFPI2, BNIP3, FOXE1, SYNE1, SOX17, PHACTR3 and JAM3, wherein detection of the epigenetic change is indicative of a predisposition to, or the incidence of, cancer. Also described are pharmacogenetic methods for determining suitable treatment regimens for cancer and methods for treating cancer patients, based around selection of the patients according to the methods of the invention. The present invention is also concerned with improved methods of collecting, processing and analyzing samples, in particular body fluid samples. These methods may be useful in diagnosing, staging or otherwise characterizing various diseases.

Abstract: Provided are methods of adding adapters to nucleic acids. The methods include combining in a reaction mixture a template ribonucleic acid (RNA), a template switch oligonucleotide including a 3? hybridization domain and a sequencing platform adapter construct, a polymerase, and dNTPs. The reaction mixture components are combined under conditions sufficient to produce a product nucleic acid that includes the template RNA and the template switch oligonucleotide each hybridized to adjacent regions of a single product nucleic acid that includes a region polymerized from the dNTPs by the polymerase. Aspects of the invention further include compositions and kits.

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: Apparatus and methods to identify nucleotides of a DNA strand. The method includes exposing the DNA strand to a first dye or peptide, attaching the first dye or peptide to a first type of nucleotide (A,T,C,G) of the DNA strand, the first dye or peptide changing a conductance of the first type of nucleotide to which the first dye or peptide is attached, and measuring a tunneling current signal for all nucleotides of the DNA strand, the changed conductance of the first type of nucleotide providing amplified tunneling current discrimination of the nucleotides of the DNA strand.

Abstract: This document provides methods and materials for assembling nucleic acid constructs (e.g., TALENs). For example, methods for assembling TALEs that are rapid, flexible for use in many cloning scaffolds (such as common nuclease and nickase backbones), and achievable with standard molecular biology laboratory tools, thereby making TALEs a more accessible genome system, are provided.

Abstract: The invention provides methods of forming a circular template for sequencing a target nucleic acid. The circular template is generated by amplification of a segment of the target nucleic acid with chimeric primers with complementary 5? ends. The circular template has a single nick or gap providing a site for initiation of template-directed extension for sequence analysis. Sequencing of a single template generates reads of alternating segments of the same strand of the target nucleic spaced by primer segments. The different reads of the same strand of the target nucleic acid can be compiled to generate a consensus sequence. Because only one strand of the target nucleic acid is sequenced per reaction, the present method avoids errors introduced by unwittingly combining sequences of both strands of a heteroduplex PCR product.

Abstract: Provided herein is a method for analyzing polynucleotides such as genomic DNA. In certain embodiments, the method comprises: (a) treating chromatin isolated from a population of cells with an insertional enzyme complex to produce tagged fragments of genomic DNA; (b) sequencing a portion of the tagged fragments to produce a plurality of sequence reads; and (c) making an epigenetic map of a region of the genome of the cells by mapping information obtained from the sequence reads to the region. A kit for performing the method is also provided.

Abstract: A composition and method for controlled in vitro fragmentation of nucleic acids. A transposase forms catalytically active complexes with a modified transposon end that contains within its end sequence degenerate, apurinic/apyrimidinic sites, nicks, or nucleotide gaps, to fragment or shear a target nucleic acid sample in a controlled process. This method yields desired average nucleic acid fragment sizes. The inventive composition and method may be applied for generation of DNA fragments containing shortened transposon end sequences to facilitate subsequent reactions, for production of asymmetrically tailed DNA fragments, etc.

Abstract: Compositions, reaction mixtures, and methods for performing an amplification reaction, including multiplex amplification reaction, wherein the method comprises using one or more amplification oligomer complexes comprising linked first and second amplification oligomer members. In one aspect, the amplification oligomer complex is hybridized to a target nucleic acid, the target nucleic acid with hybridized amplification oligomer complex is then captured, and other components are washed away. Target sequences of the target nucleic acids are pre-amplified to generate a first amplification product. The first amplification product is amplified in one or more secondary amplification reactions to generate second amplification products.

Abstract: The invention relates to methods of generating templates for a nucleic acid sequencing reaction which comprise: providing at least one double-stranded nucleic acid molecule, wherein both strands of the double-stranded nucleic acid molecule are attached to a solid support at the 5? end, cleaving one or both strands of the double-stranded nucleic acid molecule, and subjecting the cleaved strand(s) to denaturing conditions to remove the portion of the cleaved strand(s) not attached to the solid support, thereby generating a partially or substantially single-stranded template for a nucleic acid sequencing reaction.

Abstract: The present technology provides for a microfluidic substrate configured to carry out PCR on a number of polynucleotide-containing samples in parallel. The substrate can be a single-layer substrate in a microfluidic cartridge. Also provided are a method of making a microfluidic cartridge comprising such a substrate. Still further disclosed are a microfluidic valve suitable for use in isolating a PCR chamber in a microfluidic substrate, and a method of making such a valve.

Abstract: The present invention relates to a recombinant microorganism comprising one or more nucleotide sequence(s) encoding: a polypeptide having ent-copalyl pyrophosphate synthase activity; a polypeptide having ent-Kaurene synthase activity; a polypeptide having ent-Kaurene oxidase activity; and a polypeptide having kaurenoic acid 13-hydroxylase activity, whereby expression of the nucleotide sequence(s) confer(s) on the microorganism the ability to produce at least steviol. The recombinant microorganism may also be capable of expressing one or more UDP-glucosyltransferases such that the microorganism is capable of producing one or more steviol glycosides.

Abstract: Methods are provided for nucleic acid analysis wherein a target nucleic acid is mixed with a dsDNA binding dye to form a mixture. Optionally, an unlabeled probe is included in the mixture. A melting curve is generated for the target nucleic acid by measuring fluorescence from the dsDNA binding dye as the mixture is heated. Dyes for use in nucleic acid analysis and methods for making dyes are also provided.

Abstract: Disclosed are a composition for PCR including polyethylene glycol-engrafted nano-sized graphene oxide (PEG-nGO), the composition for PCR being capable of increasing the efficiency and specificity of PCR and shortening PCR time, and a PCR method using the same.

Abstract: The disclosure relates to a fluorescence detection instrument, including a base, a heating module, a detecting module, an illumination module, and an actuation module. The heating module, the detecting module, and the actuation module are disposed on the base. The heating module includes plural heating holders, wherein each of the plural heating holders is adapted to accommodate a light-transmissive reaction container adapted to contain a fluorescent reaction mixture with at least one targeted fluorescent probe respectively. The detecting module is configured with the heating module to form plural detection channels, wherein the plural heating holders are located at the plural detection channels respectively. The actuation module is connected with the illumination module and adapted to drive the illumination module to move to at least one predetermined position to selectively match at least one combination of the heating holder on the corresponding detection channel.