Abstract: Provided herein are methods for inducing CRISPR/Cas-based gene regulation (e.g., genome editing or gene expression) of a target nucleic acid (e.g., target DNA or target RNA) in a cell. The methods include using modified single guide RNAs (sgRNAs) that enhance gene regulation of the target nucleic acid in a primary cell for use in ex vivo therapy or in a cell in a subject for use in in vivo therapy. Additionally, provided herein are methods for preventing or treating a genetic disease in a subject by administering a sufficient amount of a modified sgRNA to correct a mutation in a target gene associated with the genetic disease.

Abstract: A method for removing an ionic liquid from an aqueous sample is provided. In some embodiments, the method includes: (a) combining an aqueous sample including an ionic liquid with an ion exchanger composition including an ion exchanger counterion to produce a solution including a fluorous salt of the ionic liquid, where at least one of the ionic liquid and the ion exchanger counterion is fluorinated; (b) contacting the solution with a fluorous affinity material, thereby removing fluorous salt from the solution and producing an aqueous eluate; and (c) collecting the aqueous eluate. In certain embodiments, the method further includes: contacting a cell with an ionic liquid composition to lyse the cell and produce an aqueous sample; and contacting the aqueous sample with a reverse phase substrate, thereby adsorbing proteins and/or lipids of the cell on the substrate. Compositions, kits and systems for practicing the subject methods are also provided.

Abstract: In a liquid chromatography system, a sample is injected into a column by flowing a solvent mixture from a mixer into the column along a solvent mixture flow path; and injecting a sample into the solvent mixture flow path downstream of the mixer. In another liquid chromatography system, a sample is injected into a column by loading an isolator fluid into a sample loop, loading a sample into the sample loop, and flowing the sample into the column as a plug in front of the isolator fluid.

Abstract: A method and a system for introducing a sample into a mobile phase of a chromatography system is provided. The method includes initially directing the mobile phase directly into a separation unit of the chromatography system, bypassing a sample loop, the mobile phase including a combined solvent, metered from a pressurized first solvent and a second solvent; loading the sample into the sample loop, while the mobile phase continues to be directed directly into the separation unit; pressurizing the sample in the sample loop with the pressurized first solvent, while the mobile phase continues to be directed directly into the separation unit; and switching the sample loop into the mobile phase, thereby introducing the pressurized sample to the separation unit.

Abstract: This invention discloses reagents and methods for increasing specificity and efficiency of RNA-guided genome editing.

Abstract: A scanner and method for using the same are disclosed. The scanner includes a stage, a MIR light source, an imaging system, and a controller. The stage is adapted to hold a specimen to be imaged and to move the specimen in a first direction and in a second direction that is orthogonal to the lateral direction. The imaging system forms an image plane of the specimen when the stage is positioned at a second direction distance, z, from a known point in the imaging system. The imaging system forms a plurality of different image planes of the specimen at the illumination wavelength. Each of the plurality of image planes is characterized by a different value of z, the controller determining a value of z for each of a plurality of points on the specimen at which the point on the specimen is in focus.

Abstract: A gas chromatographic (GC) unit or module may include one or more microfluidic devices, a GC column, and a flow controller (FC) comprising an FC input port for controlling fluid flows and pressures. The GC unit may be reconfigurable to provide different functionalities. The GC unit may be fluidly coupled to various other fluidic devices, such as other GC units, sample inlets, GC detectors, and the like. Multiple GC units and other fluidic devices may be utilized to build GC devices and associated systems of flexible, reconfigurable, and scalable architecture, thereby enabling a variety of modes of operation useful for present and future GC method development.

Abstract: An electrode assembly is provided in a high sub-atmospheric pressure region of an ion source, between an ionization chamber and a vacuum region of a spectrometer, such as a mass spectrometer, an ion mobility spectrometer, or an ion mobility-mass spectrometer. The electrode assembly is spaced at a distance from an outlet of an ion transfer device. A voltage source imparts a potential difference between the ion transfer device and the electrode assembly to accelerate ions emitted from the outlet to a collision energy. The collision energy is effective to cause collisional heating of ions in the high sub-atmospheric pressure region without voltage breakdown. The collision energy may be set to cause unfolding of folded biomolecular ions and/or dissociation of ions.

Abstract: A time-of-flight mass spectrometer (TOF-MS) utilizes a multi-channel ion detector to detect ions traveling in separate flight paths, spatially dispersed along a drift axis and/or a transverse axis, in a flight tube of a TOF analyzer. The ion beams may be dispersed by drift energy, deflection along the drift and/or transverse axis, ion mass, or a combination of two or more of the foregoing. The dispersion may be carried out before, at, or after an ion accelerator of the TOF analyzer. Ion packets may be accelerated into the flight tube at a multi-pulse firing rate. Tandem MS may be implemented on parallel ion beams simultaneously.

Abstract: An apparatus that facilitates the manual sorting of objects is disclosed. The apparatus includes a display surface having a surface that can be selectively illuminated and that is adapted for receiving the objects. An identification reader reads identification information stored on the objects. A controller causes an area on the display surface corresponding to one of the objects to be illuminated based on the identification information and indicates a location to which the illuminated object is to be moved. In one aspect of the invention, the display surface includes an area adapted for positioning a receiver for the objects. The display surface provides an indication of a position in the receiver at which the one of the objects is to be placed.

Abstract: Systems and methods for displaying gene- and/or protein-related data with respect to chromosome maps at locations identifying relevant positioning of the genes with which the gene- and/or protein related data are associated. Multiple experiments may be plotted onto the display adjacent one or more chromosome maps. Automatic extraction of genomic location, based on accession numbers or other unique identifiers and cross connection with expression data is provided. Statistical assessments of correlations between expression and genome localization may be performed. Zooming capabilities, thumbnail/fullview toggling, browsability and linked data may be included as features of the visualization systems described.

Abstract: A method of deprotecting a solid support bound polynucleotide includes the step of contacting the polynucleotide with a composition comprising a diamine under conditions sufficient to deprotect the 2?-protected ribonucleotide residue. The solid support bound polynucleotide has at least one 2?-protected ribonucleotide residue, which has the following structure: wherein BP is a protected or unprotected heterocycle; R12 is a protecting group selected from a hydrocarbyl, a substituted hydrocarbyl, an aryl, and a substituted aryl; X is O or S; and PG is a thionocarbamate protecting group.

Abstract: Among other things, this disclosure provides a method of detecting a target nucleic acid. Aspects of the method include: (a) obtaining a labeled nucleic acid probe that is complementary to a target nucleic acid, wherein the probe comprises a capture tag; (b) hybridizing the probe with the target nucleic in a fixed cell, in situ, to produce a duplex; (c) linking the probe in the duplex to a peroxidase conjugate via the capture tag to produce a peroxidase-labeled duplex; and (d) incubating the peroxidase-labeled duplex with a peroxidase substrate, wherein the peroxidase activity of the peroxidase conjugate catalyzes deposition of the substrate in the vicinity of the duplex, thereby producing a detectable signal.

Abstract: A fluidic device includes a planar structure constituted by a plurality of laminated layers and accommodating a fluid channel extending up to a surface of the planar structure, and a female adapter piece configured for a fluid-tight accommodation of a male adapter piece having a fluid conduit. The the female adapter piece is connected or connectable with the planar structure so that, when the male adapter piece is accommodated in the female adapter piece, the fluid conduit is brought in fluid-tight fluid communication with the fluid channel. The fluid channel is exposed to the female adapter piece at a lateral surface of the planar structure at which the laminated layers are exposed.

Abstract: Methods of conjugating oligonucleotides are provided. The methods may include: activating a terminal of first oligonucleotide using a squarate reagent to produce an activated first oligonucleotide; and binding the first oligonucleotide and a second oligonucleotide to a splint oligonucleotide; to conjugate the activated first oligonucleotide with a terminal of the second oligonucleotide via a squaramide linkage to produce a squaramide-linked oligonucleotide. Also provided are oligonucleotides that include a squaramide internucleoside linkage. Compositions are provided that include a first oligonucleotide including 300 or more nucleosides and at least one squaramide internucleoside linkage; and a second complementary oligonucleotide not including a squaramide linkage. Kits and compositions for practicing the subject methods are also provided.

Abstract: Apparatus for carrying out spatially offset Raman spectroscopy (SORS) is described. The apparatus comprises a rotatable prism arranged such that a spatial offset between an entry region and a collection region at a sample is dependent upon an angle of rotation of the prism.

Abstract: A light source having a gain chip, a retro reflecting prism and a first actuator is disclosed. The gain chip amplifies light passing therethrough. The retro reflecting prism is characterized by a pivot axis within the retro reflecting prism, an input light direction, an output light direction, and a diffraction grating that receives light emitted by the gain chip traveling in the input direction, returns a diffracted light beam to the gain chip along the input light direction and generates an output light beam. The first actuator causes the retro reflecting prism to rotate about the pivot axis in response to a control signal being coupled to the first actuator.

Abstract: An ion source includes a plasma generator for supplying plasma at an ionization region proximate to a sample surface. The plasma generator applies energy that may be utilized for desorbing analytes from the sample surface as well as for generating plasma by which analytes are excited or ionized. Desorption and ionization/excitation may be controlled as individual modes. The ion source may be interfaced with an ion-based or optical-based spectrometer. A sample support may be provided, which may be capable of performing analytical separation.

Abstract: A method for fragmenting a genome is provided. In certain embodiments, the method comprises: (a) combining a genomic sample containing genomic DNA with a plurality of Cas9-gRNA complexes, wherein the Cas9-gRNA complexes comprise a Cas9 protein and a set of at least 10 Cas9-associated guide RNAs that are complementary to different, pre-defined, sites in a genome, to produce a reaction mixture; and (b) incubating the reaction mixture to produce at least 5 fragments of the genomic DNA. Also provided is a composition comprising at least 100 Cas9-associated guide RNAs that are each complementary to a different, pre-defined, site in a genome. Kits for performing the method are also provided. In addition, other methods, compositions and kits for manipulating nucleic acids are also provided.