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This protocol describes the establishment of a reversible replication barrier using plasmid templates containing a lacO array bound by LacR repressor. The method allows fine control of replication fork movement and replication fork encounter with DNA lesions.
It can be challenging to obtain meaningful and accurate structural information for air-sensitive proteins. This protocol describes the application of customized vacuum manifold and anaerobic chamber setups for the purification and cryo-electron microscopy analysis of air-sensitive nitrogenase enzymes.
Attempts to reproduce the computational steps described in published omics research often fail. This review provides guidelines for the packaging and containerization of software so that readers can use the exact programs used in published work.
Isotopically labeled amino acids are useful in pharmacology and for medical imaging. In this protocol, C1-labeled α-amino acids are prepared via late-stage carboxylate exchange of unprotected α-amino acids with [*C]CO2 where *C is 13C, 11C or 14C.
We present a protocol for achieving efficient generation of hPSC-CM aggregates in suspension culture, emphasizing process simplicity, robustness and GMP compliance. The strategy promotes clinical translation and other applications that require large numbers of CMs.
This protocol can be used to generate three-dimensional vascularized bone marrow organoids from human induced pluripotent stem cells. The organoids contain key stromal and hematopoietic cell types and can be engrafted with normal and malignant cells from adult donors to model niche interactions.
Cell engineering using polymeric material is an area that remains largely unexplored. This protocol describes two light-driven approaches for synthesizing bioactive polymers within intricate intracellular settings.
This protocol is for using PanSyn, the first software package for the identification of micro- and macrosynteny and their functional integration for comprehensive characterization of genome architecture and regulatory evolution.
Genetic interactions have been found to influence phenotypes in a variety of systems, yet their specific contribution to complex diseases remains unclear. This protocol describes Bridging Gene sets with Epistasis (BridGE), a computational approach for discovering interactions between biological pathways from genome-wide association studies data.
The protocol presents for an optimized culture system for deriving porcine expanded potential stem cells from preimplantation embryos and reprogrammed somatic cells, and for validation and characterization.
This protocol presents a method for single-cell RNA sequencing of tissue-resident murine eosinophils, with a complementary method for CRISPR screening of bone marrow-derived eosinophils.
BEHAV3D is a 3D live imaging platform for analyzing engineered T cell behavior and tumor response. This provides insights into the mode of action of cellular immunotherapy, capturing heterogeneity within and between tumors during treatment response.
MXenes are two-dimensional materials with diverse optoelectronic, biological, mechanical and chemical properties. This protocol describes how to prepare single-layer flakes of Ti3C2Tx, the most important and widely used MXene, from a Ti3AlC2 MAX phase precursor.
Synteny and colinearity are important parameters that delineate the evolution of genomes and gene families. This protocol describes MCScanX, a user-friendly toolkit that facilitates rapid evolutionary analysis of chromosomal structural changes.
Transkingdom Network Analysis (TkNA) is a unique analytical framework for inferring causal factors underlying host–microbiota and other multi-omic interactions, by integrating data from multiple cohorts and diverse omics types.
Ultra-stable magnetic tweezers allow measuring individual protein dynamics in equilibrium under physiologically relevant pulling forces and over timescales of days to weeks, enabling high-precision molecular studies in mechanobiology.
Atomic force microscopy can be used to determine the stiffness of materials. This protocol describes how to measure and quantify the Young’s modulus E of pulmonary mouse and human basement membranes with atomic force microscopy and the Center for Applied Tissue Engineering and Regenerative Medicine processing toolbox.
We provide a twisting fabrication process for fiber electrodes that can be assembled into electronic threads and then integrated in electronic textile-based wearables.