Volume 12 Issue 9, September 2017

This protocol extension describes how to obtain functionally mature oocytes from embryonic or induced pluripotent stem cells. These oocytes can be used to produce live mouse offspring.

The lipid cubic phase (in meso) method is used for generating crystals and X-ray structures of integral membrane proteins. This protocol describes the cubicon method for concentrating membrane proteins in the cubic mesophase.

Cheng & Westerblad describe the mechanical dissection of living single fibers from the mouse flexor digitorum brevis muscle. Isolated intact fibers are subsequently used for force and myoplasmic free [Ca²⁺] measurements.

This protocol describes how to use sedimentation velocity analytical ultracentrifugation in combination with fluorescence optical detection for the analysis of mass, shape, size distribution, and binding constants of interacting proteins.

This protocol describes procedures for using the genetic tag APEX2 to generate contrast for electron microscopy in cultured cells.

This protocol describes how to obtain functionally mature oocytes from primordial germ cells from fetal mouse ovaries in vitro. These oocytes can be used to produce live mouse offspring.

This protocol describes a phagemid-based intracellular evolution approach to generate and select for proteins with improved biological characteristics.

The Technau lab provides their protocol for the generation of stably transgenic sea anemones. An expression vector is digested with the meganuclease I-SceI and then microinjected into embryos, where I-SceI mediates stable integration into the genome.

This protocol describes how to measure telomere length in archival human cardiac tissues using cardiac quantitative fluorescent in situ hybridization (CQ-FISH) in a cell-type-specific manner.

This protocol describes how to exert precise spatial and mechanical control over genetically encoded cell-surface receptors in live cells using magnetoplasmonic nanoparticles.

This protocol differentiates hPSCs into self-renewing epicardial cells by appropriate differentiation-stage-specific application of Gsk3 inhibitor, Wnt inhibitor, and then Gsk3 inhibitor again in a completely defined, xeno-free system.

Aerolysin nanopores are being used to discriminate between oligonucleotides of different length, composition and concentration. This protocol describes the procedures for aerolysin nanopore formation in lipid bilayers, quality checks and data analysis.

Titration of the optimal ratios of multicomponent biochemical reactions can be a laborious task. This protocol describes how to explore ∼10,000 combinations of concentrations of one to three reaction components with a droplet-based microfluidics platform.

This protocol describes how to detect protein–DNA and protein–RNA interactions in plant leaves using a FRET–FLIM approach.

This protocol describes how to investigate genome-wide or locus-specific chromatin ADP-ribosylation using ADPr-ChAP.

Ventral midbrain dopamine progenitors are obtained from human pluripotent stem cells using a fully defined and xeno-free differentiation protocol. The progenitors can be cryopreserved or used for transplantation or terminal maturation in vitro.

This protocol describes an optimized procedure for retroviral (RV) transduction of mouse T cells by enriching RV-susceptible activated CD8⁺ T cells through Percoll density centrifugation, enhancing the frequency of RV-transduced cells ∼sixfold in vivo.

This protocol describes how to design and produce hierarchically assembled nanomaterials with tunable functionalities using engineered M13 bacteriophages.

This protocol monitors how fusion of proteoliposomes containing the vesicle-associated SNARE synaptobrevin with proteoliposomes containing the target-associated SNAREs syntaxin-1 and SNAP-25 depends on Munc18-1, Munc13-1, synaptotagmin-1, NSF and α-SNAP.