Volume 14 Issue 8, August 2019

Selective ribosome profiling (SeRP) reveals nascent chain length–resolved binding profiles of a co-translationally acting factor and relies on selective enrichment of factor-engaged monosomes. This protocol describes how to perform the procedure in yeast.

Cross-linking with mass spectrometry (XL-MS) can reveal the topology of protein complexes. This protocol describes how to synthesize a cleavable cross-linker and use it to map protein structures and interactions within intact cells and animal tissues.

This protocol details labeling of bacterial outer-membrane proteins with spin labels to study conformational changes and their interaction with ligands and substrates in situ, using pulsed electron–electron double resonance (PELDOR or DEER) spectroscopy.

A protocol for the design, construction, and operation of an intelligent image-activated cell sorting (iIACS) machine that performs real-time image-based sorting of single cells from heterogeneous populations with high throughput and intelligence.

DNA origami can be used to create a wide diversity of 3D structures. This protocol describes a DNA origami silicification (DOS) approach for generating complex silica composite nanomaterials with tailored structural properties.

This protocol describes the design of a paper-based analytical device with printed electrodes where the reagents are incorporated into the paper. Such a device can be used in conjunction with a portable detector connected to a PC or smartphone.

This protocol describes procedures for generating genome-edited mouse models by injecting CRISPR reagents into oviducts of pregnant females and subsequently electroporating the reagents into zygotes in situ, thus bypassing cumbersome ex vivo handling of embryos.

The transmembrane domains of many type I/II membrane proteins oligomerize in the lipid bilayer, mediating protein assembly and clustering that are critical to their function. The authors provide a general protocol for determining the structures of these domains in a near–lipid bilayer environment.

This protocol describes how to create functional chromosome fusions in yeast through sequential rounds of CRISPR–Cas9-guided homologous recombination. Each round of pairwise chromosome end-to-end fusion deletes two telomeres and one centromere.

Whole-cell extracts are analyzed by gas chromatography–mass spectrometry to monitor interactions among all enzymes of distal cholesterol biosynthesis in a single experiment for inhibitor screening.

Here, the authors describe rhTCRseq, RNase H–dependent PCR-enabled TCR sequencing, for repertoire analysis from bulk RNA samples or single-cell profiling.