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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 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 flow cytometric identification of viral translation-competent reservoirs, based on concurrent detection of cellular HIV Gagpol mRNA by in situ RNA hybridization combined with antibody staining for the HIV Gag protein.
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.
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 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.
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.
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.
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.
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.
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.
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 a phagemid-based intracellular evolution approach to generate and select for proteins with improved biological characteristics.