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The authors describe a process for the mass production and cryopreservation of definitive endodermal cells derived from human pluripotent stem cells using a chemically defined, xeno-free suspension culture in stirred tank bioreactors or rotating Erlenmeyer flasks.
This protocol enables the sensitive detection of norovirus from environmental water samples in situ using a custom-built smartphone-based fluorescence microscope to detect anti-norovirus antibody-conjugated fluorescent particles on a paper microfluidic chip.
This protocol describes how to successfully design and execute molecular brightness analysis of G-protein–coupled receptor oligomerization. Labeling strategies, controls and instructions for spatial and temporal brightness imaging and data analysis are provided.
The authors describe a reverse genetic system that enables rapid synthesis of wild-type, mutant and reporter SARS-CoV-2 strains to study viral infection, transmission, pathogenesis, therapeutics and vaccines.
This protocol describes the generation, maintenance and applications of snake venom gland organoids. These organoids can be derived within days from embryonic or adult venom gland tissues from various snake species.
This protocol describes strategies for making N-heterocyclic carbene gold(I) chloride complexes. These can be used as synthons to access various oxygen-, nitrogen- or carbon-bound gold complexes that are widely used as catalysts.
This protocol describes procedures for isolation of nuclei of multiple cell populations (neurons, microglia, oligodendrocytes, and astrocytes) from resected or postmortem brain tissue that are compatible with downstream epigenomic or transcriptomic profiling.
This protocol describes how to isolate up to six different subpopulations of extracellular vesicles (EVs) from tissues. The procedure includes detailed instructions for EV characterization using electron microscopy, RNA and protein analysis.
This protocol describes how to use natural language processing software to analyze metabolomics data to prioritize metabolites for further study, identify candidates for unique disease biomarkers and elucidate their function on a pathway level.
This protocol describes a cell-free method for experimentally identifying genome-wide off-target sites of CRISPR nucleases and deaminases through in vitro digestion of genomic DNA or chromatin followed by whole-genome sequencing.
This protocol describes procedures for high-throughput analysis of trigenic interactions in yeast. Triple-mutant strains generated in a series of automated replica-pinning steps are grown on agar plates as individual colonies, and interactions are quantified with the trigenic synthetic genetic array scoring method.
This protocol describes how to perform near-infrared spectroscopy and imaging of connective tissues. Detailed guidelines are provided for sample preparation, spectral acquisition and data pre-processing and analysis, with example applications.
The authors describe a modular pipeline to detect aberrant gene expression events (expression level, splicing and mono-allelic expression) from patient RNA sequencing data, which can complement DNA-based diagnosis by enhancing the functional interpretation of variants.
This protocol for base editing in cultured mammalian cells provides guidelines for choosing target sites, appropriate base editor variants and delivery strategies, as well as detailing the computational analysis of base-editing outcomes using CRISPResso2.
This protocol describes how to perform combined protein isoform detection with nucleic acid analysis from the same individual mouse embryo or blastomere using fractionation polyacrylamide gel electrophoresis (fPAGE).
A protocol for implementing Brillouin microscopy to study biological materials. The procedure contains instructions for integrating an add-on Brillouin module with an existing confocal microscope as well as for its calibration and use in data collection and processing.
Reversible protection of the primary face of cyclodextrins by silylation is a very popular strategy for modification of the secondary rim. This protocol describes how to prepare these important intermediates in high yield and purity.
Cultivating native bacteria from fresh plant roots is essential for understanding their interaction with the host plant. This protocol describes their isolation and accurate taxonomical identification using two-sided barcode polymerase chain reaction and Illumina sequencing.
The authors describe hardware setup and experimental workflows for collecting and analyzing the biotic and abiotic environmental exposome at the individual level.