Volume 17 Issue 11, November 2022

This protocol describes prime editing (PE) and twinPE experiments as well as the design and optimization of pegRNAs. The authors provide guidelines for selecting the proper PE system for a given application and how to perform PE in mammalian cells.

Small-animal blood exchange replaces or dilutes blood components more precisely than parabiosis does. This protocol describes the exchange process, provides blueprints of the required device and derives equations for simulating dilution of biomolecules.

Photochemical methods are increasingly being used in chemical synthesis. This protocol describes how to optimize the reaction conditions for a standard chiral example using cyclodextrin derivatives as supramolecular hosts.

This protocol describes the isolation from brain tissue of extracellular vesicle subpopulations, including microvesicles, exosomes and mitochondria-derived mitovesicles, using a high-resolution (iodixanol) density step gradient. Extracellular vesicle characterization and analysis are also presented.

Volcano plots and activity maps are powerful tools for studying homogeneous catalysis. This protocol describes how to build these from computed DFT data and use them to predict and rationalize the performance of homogeneous catalysts.

This protocol provides a step-by-step guide to building an affordable single-molecule localization microscopy setup for high-throughput super-resolution imaging by using off-the-shelf and machined parts.

This protocol includes a suite of procedures for measuring the passive permeability of solutes through membranes using fluorescence-based assays that report either volume or pH changes of synthetic vesicles or live cells.

Contamination of water by oil is a problem both in the environment and in engineering applications. This protocol describes how to prepare a transparent superoleophobic film that can coat lenses or make separation membranes for use underwater.

Identifying metabolites in cells of the tumor microenvironment is an area of intense study. This protocol describes specimen collection, enrichment of human immune cell populations and analysis by flow cytometry and LC–MS.

This protocol uses signaling networks and molecular markers recently elucidated from single-cell transcriptomic analysis of mouse foregut organogenesis to differentiate human pluripotent stem cells into digestive and respiratory organ-specific mesenchyme.