Disclosed herein include systems, methods, compositions, and kits for labeling nucleic acid targets in a sample. In some embodiments, nucleic acid targets (e.g., mRNAs) are initially barcoded on the 3′ end and are subsequently barcoded on the 5′ end following a template switching reaction and intermolecular and/or intramolecular hybridization and extension. 5′- and/or 3′-barcoded nucleic acid targets can serve as templates for amplification reactions and/or random priming and extension reactions to generate a sequencing library. The method can comprise generating a full-length sequence of the nucleic acid target by aligning a plurality of sequencing reads. Immune repertoire profiling methods are also provided in some embodiments.

Disclosed herein are method for separating, amplifying, or detecting a nucleic acid from a sample may comprise contacting a sample lysate with a plurality of buoyant, inorganic, nucleic-acid-capture microspheres. The nucleic-acid-capture microspheres may comprise unicellular hollow microspheres having a diameter between 5 and 300 μm and/or a true particle density between 0.05 and 0.60 grams/cm.sup.3. The microspheres may comprise hollow soda-lime-borosilicate microspheres. In some embodiments, the microspheres comprises hollow soda-lime-borosilicate microspheres surrounded by an amorphous silica shell. Also disclosed are kits for performing the methods.

Disclosed herein are method for separating, amplifying, or detecting a nucleic acid from a sample may comprise contacting a sample lysate with a plurality of buoyant, inorganic, nucleic-acid-capture microspheres. The nucleic-acid-capture microspheres may comprise unicellular hollow microspheres having a diameter between 5 and 300 μm and/or a true particle density between 0.05 and 0.60 grams/cm.sup.3. The microspheres may comprise hollow soda-lime-borosilicate microspheres. In some embodiments, the microspheres comprises hollow soda-lime-borosilicate microspheres surrounded by an amorphous silica shell. Also disclosed are kits for performing the methods.

Embodiments of the instant disclosure relate to novel compositions and methods for analyte detection by quantifying transcription of RNA using CRISPR-based approaches. In certain embodiments, constructs of use in assays disclosed herein can include a novel double-stranded DNA sequence having at least one transcriptional promoter, at least one regulatory element, and a target sequence. In some embodiments, constructs disclosed herein can be used in a system for quantifying transcriptional output in a sample in order to detect presence of an analyte. In other embodiments, constructs and systems disclosed herein can be used to measure enzyme activity, to detect and/or quantify at least one agent that modulates transcription, detect and/or quantify analytes (e.g. a contaminant, biomarkers or other agent) or a combination thereof in accordance with assessing health of a subject or assessing environmental conditions or contaminants.

Embodiments of the instant disclosure relate to novel compositions and methods for analyte detection by quantifying transcription of RNA using CRISPR-based approaches. In certain embodiments, constructs of use in assays disclosed herein can include a novel double-stranded DNA sequence having at least one transcriptional promoter, at least one regulatory element, and a target sequence. In some embodiments, constructs disclosed herein can be used in a system for quantifying transcriptional output in a sample in order to detect presence of an analyte. In other embodiments, constructs and systems disclosed herein can be used to measure enzyme activity, to detect and/or quantify at least one agent that modulates transcription, detect and/or quantify analytes (e.g. a contaminant, biomarkers or other agent) or a combination thereof in accordance with assessing health of a subject or assessing environmental conditions or contaminants.

The present application provides imidazolyl pyrimidinylamine inhibitors of cyclin-dependent kinase 2 (CDK2), as well as pharmaceutical compositions thereof, and methods of treating cancer using the same.

Some embodiments presented in this disclosure concern an Automated Tissue Dissection (ATD) System. An ATD system is a one stop, and potentially low-cost, system to perform dissections on a substrate from pathologist digital mark or pen mark on the substrate using non-contact and/or mechanical method to extract a Formalin-Fixed Paraffin-Embedded (FFPE) tissue sample with: (a) only the ROI or ROIs as area to be saved; and (b) remove or decompose nucleic acid content in the region of no interest (RONI) and collect all tissue sample from a standard microscope substrate into a specific container.

Some embodiments presented in this disclosure concern an Automated Tissue Dissection (ATD) System. An ATD system is a one stop, and potentially low-cost, system to perform dissections on a substrate from pathologist digital mark or pen mark on the substrate using non-contact and/or mechanical method to extract a Formalin-Fixed Paraffin-Embedded (FFPE) tissue sample with: (a) only the ROI or ROIs as area to be saved; and (b) remove or decompose nucleic acid content in the region of no interest (RONI) and collect all tissue sample from a standard microscope substrate into a specific container.

A next generation DNA sequencing method and use for accurate and massively parallel quantification of one or more nucleic acid targets. More particularly, the present disclosure is related to the method and a kit comprising probes for detecting and quantifying genetic targets in complex DNA pools primarily used for genetic target and variant detection in human and animal populations and environmental samples. Furthermore, the present disclosure finds particular application in the field of detection of disease-causing genetic alterations in samples obtained from human body, including without limiting biopsies, saliva and other secretions, exhaled moisture extracts, tissue, blood plasma (liquid biopsies) or the like. The present disclosure includes one or more target-specific nucleic acid probes per genetic target (left probe and right probe) and a bridge oligo.

A next generation DNA sequencing method and use for accurate and massively parallel quantification of one or more nucleic acid targets. More particularly, the present disclosure is related to the method and a kit comprising probes for detecting and quantifying genetic targets in complex DNA pools primarily used for genetic target and variant detection in human and animal populations and environmental samples. Furthermore, the present disclosure finds particular application in the field of detection of disease-causing genetic alterations in samples obtained from human body, including without limiting biopsies, saliva and other secretions, exhaled moisture extracts, tissue, blood plasma (liquid biopsies) or the like. The present disclosure includes one or more target-specific nucleic acid probes per genetic target (left probe and right probe) and a bridge oligo.