Abstract: This invention relates to the storage on a solid matrix of genetic material, in particular DNA that has been purified prior to the application to the solid matrix. More specifically, the invention relates to a solid matrix for the storage of purified DNA, which matrix has been treated with a solution comprising plant polysaccharide inulin. One advantage of the invention is that an increased amount of DNA can be stored in the solid matrix of the present invention.

Abstract: The present disclosure provides methods for determining the status of an allograft within a transplant recipient from genotypic data measured from a mixed sample of DNA comprising DNA from both the transplant recipient and from the donor. 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 invention provides, among other things, methods of purifying messenger RNA (mRNA) including the steps of subjecting an impure preparation comprising in vitro synthesized rnRNA to a denaturing condition, and purifying the rnRNA from the impure preparation from step (a) by tangential flow filtration, wherein the mRNA purified from step (b) is substantially free of prematurely aborted RNA sequences and/or enzyme reagents used in in vitro synthesis.

Abstract: A method of single-stranded RNA purification comprising the steps applying a sample containing single-stranded RNA to a solid phase bearing dominantly or exclusively primary amino groups on its surface, at a pH value sufficient to bind at least predominantly the single-stranded RNA, eluting the single-stranded RNA from the surface of the solid phase by exposing the surface of the solid phase to increasing pH.

Abstract: The present invention relates to method for producing and purifying RNA comprising the steps of providing DNA encoding the RNA; transcription of the DNA into RNA; and conditioning and/or purifying of the solution comprising transcribed RNA by one or more steps of tangential flow filtration (TFF).

Abstract: A system, composition, method and kit for removing a detergent, such as an anionic detergent, from an aqueous solution, comprising a salt, a water immiscible alcohol of Formula I: R1—OH??(Formula I) where R1 is an optionally substituted, linear, branched or cyclic C4-C12 alkyl; and a water immiscible halocarbon, wherein said halocarbon is miscible with said alcohol of Formula I. The system can be used on aqueous solutions that contain detergents (such as Sodium Dodecyl Sulphate (SDS), for example), and any detergent-associated or detergent-bound molecules that may be present in the aqueous solution, to form an aqueous phase and a non-aqueous phase, for effectively removing the detergent and any detergent-associated or detergent-bound molecules, and sequestering them into the non-aqueous phase.

Abstract: The present disclosure provides methods and compositions for the treatment of diseases and genetic disorders linked to SURF1 loss and/or misfunction. The methods and compositions of the present disclosure comprise rAAV vectors and rAAV viral vectors comprising transgene nucleic acid molecules comprising nucleic acid sequences encoding for a SURF1 polypeptide.

Abstract: Methods and materials are disclosed for use in recovering a biopolymer from a solution. In particular, the invention provides methods for extraction and isolation of nucleic acids from biological materials. The nucleic acids can be separated by forming a stable complex with soluble polysaccharide polymers and magnetic particles, in the presence of detergents and solvent. When the particles are magnetically separated out of the solution, the nucleic acids are separated with them. The nucleic acids can subsequently be released and separated from the particles. The nucleic acid preparation is useful for achieving efficient and accurate results in downstream molecular techniques such as quantification, identification of the source of the nucleic acids, and genotyping.

Abstract: Provided herein are methods of purifying messenger RNA (mRNA) by subjecting a preparation comprising in vitro synthesized mRNA to one or more steps of enzymatic digestion with a proteinase, optionally with a further oligo dT affinity chromatography step. Also provided are mRNA purified by the methods described herein.

Abstract: Provided herein is a system comprising: a) spatial sampler system configured to collect and transmit one or a plurality of cells or nuclei from a tissue specimen, the system comprising: i) a lower carrier having an array of conduits passing therethrough, each conduit comprising an opening on a first side and communicating with an opening on a second side, wherein the opening on the second side either (1) terminates in a capillary or (2) opens onto a well of a multiwell plate; ii) positioned, above the lower carrier, a perforated specimen holder configured to support a frozen tissue specimen, wherein the specimen holder comprises a plurality of perforations having a size sufficient to permit the passage of single cells or nuclei; and iii) positioned, above the specimen holder, a multifunctional head comprising an upper array of upper conduits, optionally, each aligned with a conduit opening of the lower carrier.

Abstract: Provided is a composition for preserving cell-free nucleic acids and/or cells in a biological sample and methods for use thereof. The composition comprises at least one volume excluding polymer, at least one osmotic agent and at least one enzyme inhibitor. The composition optionally further comprises at least one metabolic inhibitor. Further, provided is a kit comprising the composition, preferably in a blood collection tube, or the components of the composition.

Abstract: The present invention provides, among other things, methods for purifying mRNA based on normal flow filtration for therapeutic use.

Abstract: Provided herein are methods and compositions to extract and enrich by, physical separation or amplification, relatively short nucleic acids from a nucleic acid composition containing a high background of longer nucleic acids (e.g., host or maternal nucleic acids; genomic nucleic acid and the like).

Abstract: The present invention relates to compositions, kits and methods for making and using RNA compositions comprising in vitro-synthesized ssRNA inducing a biological or biochemical effect in a mammalian cell or organism into which the RNA composition is repeatedly or continuously introduced. In certain embodiments, the invention provides compositions and methods for changing the state of differentiation or phenotype of a human or other vertebrate cell. For example, the present invention provides mRNA and methods for reprogramming cells that exhibit a first differentiated state or phenotype to cells that exhibit a second differentiated state or phenotype, such as to reprogram human somatic cells to pluripotent stem cells.

Abstract: The present invention relates to a new apparatus and a new method for the purification and the recovery of extra-chromosomal nucleic acids sequence(s).

Abstract: The invention relates to a new process for the purification of oligonucleotides which comprises the removal of the acid labile 5?hydroxy protecting group at the 5?-O-oligonucleotide terminus of the oligonucleotide by way of tangential flow filtration with an acidic buffer solution. The process requires less steps and allows a higher degree of automation.

Abstract: Disclosed herein, inter alia, are compositions and methods for depleting nucleotide impurities in nucleotide solutions.

Abstract: Method of analysis for alleles of a target. In the method, a plurality of fluid volumes may be formed. Each fluid volume may contain a first primer pair to amplify a first allele of a target, a second primer pair to amplify a second allele of the target, a first reporter including a first photoluminophore and providing a primer of the first primer pair, and a second reporter including a second photoluminophore and providing a primer of the second primer pair. Each of the first and second reporters may include an oligomer having a quencher, and the oligomer may be configured to base-pair with the primer of the first primer pair and the primer of the second primer pair. The first and second alleles may be amplified using the first and second primer pairs. Photoluminescence may be detected. A level of each allele may be determined.

Abstract: The present disclosure relates to a composition and its use for treating a sputum sample suspected to contain mycobacteria. The composition comprises thymol, a linear or branched alcohol, a chaotropic agent, a reducing agent, a detergent, and a buffer, and has a pH value between 8.5 and 10. Also disclosed is a method for treating a sputum sample suspected to contain mycobacteria.

Abstract: The present disclosure provides, among other things, a recombinant adeno-associated virus (rAAV) vector comprising an AAV8 or AAV9 capsid and a codon-optimized sequence encoding a human iduronate-2-sulfatase (I2S) enzyme. The disclosure also provides a method of treating a subject having Hunter syndrome (MPS II), comprising administering to the subject in need thereof a recombinant adeno-associated virus (rAAV) vector comprising an AAV8 or AAV9 capsid, and a promoter operably linked to a nucleic acid sequence that encodes iduronate-2-sulfatase (I2S), and wherein administering results in an increase in I2S enzymatic activity in the subject.

Abstract: Disclosed herein are embodiments of a fixed magnet assembly that can be implemented into automated platforms for performing polynucleotide extraction from biological samples and preparing the polynucleotides into an amplification-ready form. The fixed magnet assembly can be used to provide magnetic energy to a container containing magnetic particles and a reaction mixture of polynucleotides, in order to bring about a separation of the magnetic particles from the reaction mixture. This can prevent or reduce magnetic particle carryover in the prepared amplification-ready sample, thereby improving amplification results.

Abstract: A disease screening device (100) comprising a substrate (101) and a sonication chamber (102) formed on the substrate (101). The sonication chamber (102) is provided with an ultrasonic transducer (105) which generates ultrasonic waves to lyse cells in a sample fluid within the sonication chamber (102). The device (100) comprises a reagent chamber (111) formed on the substrate (101) for receiving a liquid PCR reagent. The device (100) comprises a controller (23) which controls the ultrasonic transducer (105) and a heating arrangement (128) which is provided on the substrate (101). The device (100) further comprises a detection apparatus which detects the presence of an infectious disease, such as COVID-19 disease.

Abstract: A gene PpHSP21 with black spot disease resistance is isolated from Pyrus pyrifolia. A nucleotide sequence of the gene PpHSP21 is shown in SEQ ID NO. 1. An amino acid sequence of an encoded protein of the PpHSP21 gene is shown in SEQ ID NO. 2. By constructing the plant overexpression vector and silencing vector of the gene PpHSP21, the gene PpHSP21 is introduced into the plant by the Agrobacterium-mediated genetic transformation method, so that the gene PpHSP21 is able to be overexpressed in the plants, thereby significantly improving black spot disease resistance in plants. The discovery and identification of the gene PpHSP21 provide new genetic resources for stress resistance molecular design and breeding in plants, and to provide new genetic resources for the implementation of green agriculture. The development and utilization of the genetic resources is conducive to reducing agricultural production costs and achieving environmental friendliness.

Abstract: Provided are methods, compositions and kits for depleting host nucleic acid in a biological sample, said sample having been previously obtained from an animal host.

Abstract: A microfluidic device for analysing a specimen comprises a loading area for loading the specimen of interest and an analytical column. The loading area is connected on two sides to a first duct and a second duct respectively, both integrated in the microfluidic device. The microfluidic device comprises a first integrated input connected to the first duct to take the specimen into the loading area, a first integrated output connected to the second duct to discharge the rest of the specimen, once it has flown through the loading area, and a second integrated output downstream the analytical column. The first integrated output is arranged for during a first loading period of time being in circuit connected to the first integrated input so as to load the sample into the loading zone of the device while preventing loss of specimen during loading of the sample into the analytical column.

Abstract: A method for analyzing planar sample is provided. In some cases the method comprises: (a) incubating the planar sample with a capture agent that is linked to an oligonucleotide, wherein the capture agent specifically binds to complementary sites in the planar sample; (b) reading a fluorescent signal caused by extension of a primer that is hybridized to the oligonucleotide, using fluorescence microscopy. Several implementations of the method, and multiplexed versions of the same, are also provided.

Abstract: Aspects of the disclosure relate to liquid chromatography (e.g., HPLC) methods which enable high resolution separations of polynucleotides having hydrophobic portions (e.g., polyadenylated nucleic acids, such as mRNA) based upon the hydrophobic character of the molecules (e.g., polyA tail length). In some embodiments, the disclosure describes liquid chromatographic methods for separating a nucleic acid having a hydrophobic portion (e.g., a polyadenylated nucleic acid, such as an mRNA) from a complex mixture by a mobile phase system that comprises an ion pairing agent selected from Tris, inorganic cations (including e.g., Na, Li, K, ammonium, etc.), biological buffers (e.g., MOPS, HEPES, PIPES, etc.), and other charged or hydrophilic moieties, and lacks conventional ion pairing agents (e.g., Triethylammonium acetate, TEAA). Accordingly, in some embodiments methods described by the disclosure are useful for assessing the quality of pharmaceutical preparations comprising nucleic acids.

Abstract: An optical gene biosensor is disclosed. The optical gene biosensor includes a substrate; a molecular beacon anchored to the substrate, wherein the molecular beacon includes an oligonucleotide specifically binding to a target nucleic acid molecule and a first compound bound to a first terminal of the oligonucleotide; an optical marker specifically binding to the first compound, wherein the optical marker is configured to retro-reflect irradiated light; a light source for irradiating the optical marker with light; and a light-receiver for receiving light retro-reflected from the optical marker. The optical gene biosensor may perform accurate quantitative analysis of a target nucleic acid molecule using both non-spectral and spectral light sources.

Abstract: According to various embodiments described herein, a microfluidics-chip based purification device and system for Sanger-sequencing reactions is provided. The device and system allow for the introduction into a sequencing system of a cartridge containing purification technologies specific to the sequencing contaminants or sequencing method where the simplified purification solution of a cartridge allows automation of the sample purification process, reduced consumption of purification reagents, and consistency in sampling by reducing the sampling errors and artifacts. These various embodiments therefore solve the need for a microfluidics-chip-based, Sanger-sequencing reaction purification system for CE devices. The microfluidic chips described can be used as a PCR chip by reorganizing the on-chip reagents, reaction wells and work flow steps.

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: Disclosed is a method for the recovery of nucleic acids from a solid support, the method comprising the steps, in any suitable order, of: a) providing a solid support at least a region of which is absorbent and impregnated with a chaotropic agent; b) combining a biological sample, possibly including nucleic acids, with the region; c) washing the region in a washing buffer solution; d) simultaneously heating and agitating the region in a further buffer solution; e) separating the region from the further buffer solution; f) extracting at least a portion of any remaining further buffer solution from the region to provide an extracted buffer solution; g) combining the further buffer solution and the extracted buffer solution portion; and h) subsequently processing the combined buffer solutions in order to amplify any nucleic acids in said combined solution. Hardware suitable for implementing the above method and a kit of parts is disclosed also.

Abstract: Compositions and methods of capturing one or more nucleic acid molecules of a cell or subcellular compartment are described. In certain aspects, the compositions comprise a caged molecule comprising one or more photolinkers and an antisense oligonucleotide, which when uncaged hybridizes to a target nucleic acid molecule.

Abstract: The present invention relates to method for producing and purifying RNA comprising the steps of providing DNA encoding the RNA; transcription of the DNA into RNA; and conditioning and/or purifying of the solution comprising transcribed RNA by one or more steps of tangential flow filtration (TFF).

Abstract: The present teachings relate to the extraction of nucleic acid from solid materials. Provided are useful compositions, methods, and kits for obtaining nucleic acids from a solid biological sample or an adhesive material having a biological material adherent or embedded within the adhesive substrate. The extracted nucleic acid can be used in downstream applications such as genotyping, detection, quantification, and identification of the source of the biological material.

Abstract: An integrated sample purification system includes a housing, a sample container rack, a filter holder, and a cylindrical magnet. The sample container rack and the filter device holder are disposed in the housing. The sample container rack is configured to hold one or more sample containers, the filter device holder is configured to hold one or more filter devices. The cylindrical magnet is adjacent to and external to the sample container rack, and is rotated about a central, longitudinal axis of the magnet by an electric motor disposed in the housing to lyse cells. Molecules of interest in the lysed cells are purified using filters that bind specifically to the molecules of interest. The system is readily amenable to automation and rapid purification and analysis of molecules of interest, such as nucleic acids and proteins.

Abstract: The present invention relates to method for producing and purifying RNA comprising the steps of providing DNA encoding the RNA; transcription of the DNA into RNA; and conditioning and/or purifying of the solution comprising transcribed RNA by one or more steps of tangential flow filtration (TFF).

Abstract: A phenol-free method for isolating a nucleic acid from a sample is provided, said method comprising the following steps: a) adding a precipitation buffer to a sample to prepare an acidic precipitation mixture wherein said precipitation buffer comprises a metal cation precipitant and a buffering agent, has a pH value of 4.0 or less and does not comprise an organic solvent selected from aprotic polar solvents and protic solvents and wherein the acidic precipitation mixture comprises the metal cation precipitant in a concentration of less than 200 mM and precipitating proteins; b) separating the precipitate from the supernatant, wherein the supernatant comprises small RNA having a length of less than 200 nt and large RNA having a length of at least 1000 nt; and c) isolating a nucleic acid from the supernatant. The present method allows to avoid the use of organic solvents during protein precipitation. Also provided is a precipitation buffer.

Abstract: This invention relates to the storage on a solid matrix of genetic material, in particular DNA that has been purified prior to the application to the solid matrix. More specifically, the invention relates to a solid matrix for the storage of purified DNA, which matrix has been treated with a solution comprising plant polysaccharide inulin. One advantage of the invention is that an increased amount of DNA can be stored in the solid matrix of the present invention.

Abstract: This invention relates to monodisperse magnetic hydrogel polymer particles comprising a magnetic material and a polymer formed from (a) a hydrophilic vinylic monomer having a log Poct/wat (log P) of less than about 0.5; and (b) a crosslinker comprising at least two vinyl groups. The invention also relates to monodisperse coated hydrogel polymer particles comprising a polymer formed from (a) a hydrophilic vinylic monomer having a log Poct/wat (log P) of less than about 0.5; and (b) a crosslinker comprising at least two vinyl groups; and a coating. Also provided are methods of forming the monodisperse magnetic hydrogel polymer particles and monodisperse coated polymer particles.

Abstract: Methods and compositions for extracting nucleic acids such as microRNAs (miRNAs) from biological samples are provided. Aspects of the methods include contacting a biological sample with proteinase K followed by contact with ferric oxide particles under acidic conditions to induce binding between the ferric oxide particles and nucleic acids (e.g., miRNAs) of the sample. In some cases, the ferric oxide particles are provided as part of a dissolvable film, which releases the ferric oxide particles upon solvation. In some embodiments, after nucleic acids bind to the ferric oxide particles, the particles are magnetically separated from the sample and are contacted with an alkaline elution buffer to release the nucleic acids.

Abstract: A disease screening device (100) comprising a substrate (101) and a sonication chamber (102) formed on the substrate (101). The sonication chamber (102) is provided with an ultrasonic transducer (105) which generates ultrasonic waves to lyse cells in a sample fluid within the sonication chamber (102). The device (100) comprises a reagent chamber (111) formed on the substrate (101) for receiving a liquid PCR reagent. The device (100) comprises a controller (23) which controls the ultrasonic transducer (105) and a heating arrangement (128) which is provided on the substrate (101). The device (100) further comprises a detection apparatus which detects the presence of an infectious disease, such as COVID-19 disease.

Abstract: There is provided a method of extracting material from a fluid method of extracting material from a fluid, the fluid being held within a fluid chamber. The method comprises drawing, with a magnetic field generating system, at least one magnetically susceptible member through the fluid around a closed path between at least three points in the chamber, said at least one member being adapted to bind to material in fluid in the chamber. The at least three points are arranged relative to each other in a shape having at least two dimensions, the magnetic field generating system being configured to move the at least on magnetically susceptible member directly between the at least three points, material in the fluid binding to the at least one magnetically susceptible member when it comes into contact with the at least one member as it moves through the fluid.

Abstract: The present invention relates to a method for lysing pathogen lysis and a method for extracting nucleic acid using zinc oxide nanostar, and the method for extracting nucleic acid using zinc oxide nanostar according to the present invention can extract nucleic acids by lysing cells of a pathogen without using a lysis buffer and can extract nucleic acid of high-purity and high-concentration by preventing nucleic acid degradation and fragmentation through various substances including salts contained in the lysis buffer at high concentration. In addition, the zinc oxide nanostar of the present invention (200 to 900 nm) has superior cell lysis capacity compared to the conventional zinc oxide nanoparticles (20 to 50 nm), thereby increasing the nucleic acid extraction efficiency and can extract at room temperature without a heating step to use as a field diagnostic method.

Abstract: A rotor assembly includes a rotor plate to rotate around a first axis, a bucket attached to the rotor plate and to rotate around a second axis, and a stop plate to rotate around the first axis between an open position and a closed position. When in the closed position, the stop plate engages the bucket to fix an angular position of the bucket relative to a plane of rotation of the rotor assembly. The rotor assembly further includes a housing for a sensor array component, the housing disposed in the bucket and including a solution inlet, a solution outlet, a transfer basin, a solution retainer disposed between the solution outlet and the transfer basin, and a collection reservoir in fluid communication with the transfer basin. The solution inlet and the solution outlet to engage ports of a flow cell of a sensor array.

Abstract: Disclosed herein are systems and methods for multiplex primer design and selection. In one example, a system includes non-transitory memory configured to store executable instructions; and a hardware processor programmed by the executable instructions to receive a plurality of target gene sequences and determine a set of primers for each target gene sequence based on a penalty score associated with the set of primers, wherein the penalty score is based on a non-linear combination of a primer-level penalty score and a set-level penalty score.

Abstract: An electrophoresis method, system, and gel recover biological substances from the gel with high efficiency. The method uses an electrophoresis gel having an injection hole into which biological substances are injected and a recovery hole from which the biological substances are recovered. The electrophoresis method includes injecting the biological substances into the injection hole, and applying an electric field penetrating the injection and recovery holes. A vertical axis in a downward direction as a positive direction is set as an X-axis, an axis which is perpendicular to the X-axis is set as a Y-axis, and coordinates of a bottom of the recovery hole are set as (XC, YC). The X coordinate XC of the bottom of the recovery hole satisfies XC>X1 when the biological substance is electrophoresed to coordinates (X1, YC) in the recovery hole from a bottom of the injection hole in the applying the electric field.

Abstract: The present disclosure relates to a method/system for preparing encapsulated exosomes from a biological sample containing exosomes, said method comprising: a) dispersing inorganic oxide particles into a buffer solution comprising a polymer and a biological sample comprising exosomes, b) allowing the polymer to react with the inorganic oxide particles to form capsules, wherein the exosomes are inside the capsules. The present disclosure also relates to a method/system for isolating and purifying encapsulated exosomes, said method comprising: a) preparing an aqueous micellar system comprising at least one surfactant, and at least one salt; b) mixing a biological sample containing encapsulated exosomes with the aqueous micellar system from step a); c) allowing the aqueous micellar system to phase separate, wherein the surfactant partitions substantially into one phase, and the other phase has a lower concentration of surfactant; and d) obtaining the encapsulated exosomes from the capsule-rich phase.

Abstract: A method of separating a target double-stranded nucleic acid molecule from a sample including the target double-stranded nucleic acid molecule and a non-target double-stranded nucleic acid molecule, including (1) mixing the sample, a pyrrole-imidazole-containing polyamide (first PI polyamide) modified with a first linker molecule and capable of specifically binding to a sequence of the target double-stranded nucleic acid molecule, and a carrier a modified with a first ligand capable of specifically binding and/or adsorbing to the first linker molecule such that a mixed solution is produced, (2) forming a complex A by binding the carrier a to the first PI polyamide with which the target double-stranded nucleic acid molecule is bound in the mixed solution, and (3) separating the complex A from the mixed solution.

Abstract: Provided herein are materials and methods for isolation of eukaryotic nucleic acid from a human or non-human animal stool sample. Also provided are methods of analysis of eukaryotic biomarkers present in a human or non-human animal stool sample.

Abstract: A collection device for a biological sample to capture target compounds such as viruses or other pathogens or particles for testing from within the sample and move the captured target compound to a separate chamber for subsequent processing. The collection device can include an openable substance blister including capture particles located in a cup interior. Capture particles can attract and bind the target compounds from the sample. An extraction tube extracts any nucleic acid from the target compound for storage or subsequent amplification and testing to confirm presence of known microorganisms. The extraction tube can comprise a heat-deformable material and can be connected to a microfluidic cartridge for further processing of nucleic acid including, amplification and detection. The microfluidic cartridge includes valves and a plurality of chambers for amplification.