Volume 12 Issue 10, October 2017

This protocol describes the isolation of gas-filled protein nanostructures, called gas vesicles, their functionalization with moieties for targeting and fluorescence, and how to use them as contrast agents for ultrasound and MRI.

This protocol describes how to use multichannel time-lapse confocal imaging of anchor-cell invasion in live Caenorhabditis elegans to monitor cell invasion through basement membranes.

This protocol describes how to generate mature adult-like cardiomyocytes by culturing mouse or human PSCs in vitro initially and then transferring to neonatal rats for further cell maturation.

Knockout Sudoku allows construction of whole-genome knockout collections for a wide range of microorganisms at a lower cost and increased speed, using combinatorial pooling, next-generation sequencing, and a Bayesian inference algorithm to process and annotate extremely large progenitor transposon insertion mutant collections.

Diazomethane is useful for inserting methyl or methylene groups in organic synthesis. Unfortunately, it is explosive. A tube-in-flask reactor where the Teflon AF-2400 tube allows only the diazomethane produced to enter the flask can be used to prepare it safely.

This protocol describes strategies for the characterization of transient protein–protein interactions and their interaction interfaces via genetically encoded releasable photo-cross-linkers.

Humanized bone-marrow-ossicle niches are formed in mice via in situ differentiation of bone-marrow-derived mesenchymal stromal cells and can be used for transplantation of normal and malignant human hematopoietic cells.

This protocol describes HyCoSuL, an approach that uses tetrapeptides containing natural and >100 unnatural amino acids to screen for protease substrate specificity and to engineer highly active and selective substrates and activity-based probes.

High-affinity magnetic immunocapture is used to rapidly purify HA-tagged mitochondria from cells for metabolite profiling. Matrix concentrations of mitochondrial metabolites are determined through LC/MS, immunoblotting, confocal microscopy, and volumetric analysis.

This protocol details the construction and use of the ichip, a platform developed to isolate previously uncultivable microorganisms from a range of environmental samples, by enabling exposure to natural growth factors through in situ culture.

This protocol describes a two-stage ‘post-translational mutagenesis’ approach for the site-specific installation of natural and unnatural amino acid side chains into recombinant proteins.