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A forebrain organoid derived from human induced pluripotent stem cells at day 52 in culture, immunostained with CTIP2 (a neuronal marker; red), PKC-λ (adherens junction marker for neural stem cells; green) and DAPI (nuclei marker; blue). Image taken from the protocol by Ming et al. doi:10.1038/nprot.2017.152. Cover design by Jamel Wooten.
This protocol extension describes DNA-free genome editing of bread wheat by delivering in vitro transcripts (IVTs) or ribonucleoprotein complexes (RNPs) of CRISPR/Cas9 by particle bombardment. The authors' previously published protocol for genome editing of wheat used CRISPR/Cas9 plasmids.
This protocol describes a cross-linking mass spectrometry (XL-MS) approach to studying homodimer interfaces, and provides procedures for stable isotope–labeling of one of the two monomers, homodimer refolding, XL-MS, and data analysis.
This protocol describes targeted chromatin capture (T2C), a high-resolution method to interrogate 3D chromatin organization and genomic interactions at sub-kilobase-pair resolution that requires minimal cell numbers and sequencing depth.
This protocol describes how to use distance restraints obtained from cross-linking mass spectrometry (XL-MS) experiments to guide the structural prediction of proteins and protein complexes.
Graphene has a unique set of extraordinary mechanical, optical, and electronic properties, but is difficult to prepare in large amounts. This protocol describes how to prepare three- to four-layer-thick graphene in stable water suspensions and powder form.
This protocol describes a set of assays for characterization of the quality and quantity of mammalian collagen type I from a diverse range of biological samples.
This protocol describes the chemical synthesis of different nonhydrolyzable diubiquitin linkages and their use in affinity purification and LC–MS/MS identification of linkage-selective diubiquitin interactors.
This protocol adapts traditional ChIP-seq for the detection of DNA secondary structures through the use of a G4-structure-specific single-chain antibody. This enables the determination of G4 structure formation genome-wide in chromatin.
This protocol describes procedures for building the SpinΩ bioreactor for 3D tissue culture and differentiating human iPSCs into different brain region–specific organoids resembling developing human dorsal forebrain, midbrain and hypothalamus.
This protocol describes an improved fluorescence lifetime imaging (FLIM) approach for nanomolar calcium imaging using the OGB-1 fluorescent calcium indicator in brain cells in situ.