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This protocol describes how to prepare and use a microfluidic device for measuring chemosensory neuronal activity in Drosophila. The device is reusable and suitable for assessing neurons in larvae, the adult proboscis and the adult leg.
Martin et al. describe a protocol to use laser microdissection to isolate specific cells or tissues from tomato fruit. This can be used to perform cell type–specific transcriptome studies in fleshy fruits, as well as in other plant tissues.
The Burgess laboratory describes their functional genomics pipeline based on CRISPR/Cas9-targeted mutagenesis in zebrafish. The system is scalable, enabling phenotypic screening of hundreds of targeted mutants.
Investigating the ramifications of site-specific protein redox modification in cells is challenging. This protocol uses HaloTagged proteins and a HaloTag-targetable photocaged 4-hydroxynonenal to elicit target-specific modifications and to trace their effects.
This protocol describes the production of Candida biofilms on the surface of polystyrene roller bottles and subsequent isolation of the biofilm extracellular matrix on a much larger scale than has previously been achieved.
This protocol describes the design and synthesis of amphiphilic gold nanocrystals coated with polymer brushes and their self-assembly into plasmonic vesicles.
This protocol from Fu et al. describes the use of self-assembled DNA nanostructures as assembly scaffolds to spatially arrange multienzyme complexes and probe their interactions.
This protocol describes how to derive haploid human embryonic stem cells (ESCs) by parthenogenesis, and how to isolate and maintain them by cell sorting. Haploid human ESCs have a wide differentiation potential and are useful for genetic screening.
This 96-well-plate ‘real-time quaking-induced conversion’ assay allows the detection of abnormal prion protein in human brain and CSF samples. It can be applied to study many protein misfolding diseases, as well as for drug screening and prion strain discrimination.
The protocol describes how to make transparent graphene neural electrodes for implantation onto the surface of the cerebral cortex in rodents and subsequent neural analysis by fluorescence microscopy, electrophysiology, optical coherence tomography, and optogenetics.
This protocol describes how to differentiate and image human embryonic stem cells on micropatterned colonies to create radially organized domains of the germ layers mimicking embryonic gastrulation in vitro.
This protocol describes how to remodel sheep and mouse fibroblast nuclei into spermatid-like structures by exogenous expression of the protamine 1 gene. Blastocyst formation following somatic cell nuclear transfer in sheep oocytes is also described.
Small-angle X-ray and neutron scattering are used to extract structural parameters and derive structural models of macromolecules in solution. The preparation of pure, monodisperse samples and exactly matching solvent blanks is crucial to the experiments' success.
This protocol extends the use of genome editing technology to the modeling or correction of large chromosomal rearrangements and short nucleotide repeat expansions. The authors use the CRISPR/Cas system to edit human induced pluripotent stem cells.
This protocol uses restriction enzymes to produce isolength tags that are concatenated and sequenced to allow genome-wide genotyping and quantification of DNA methylation levels.
Electron cryo-tomography produces 3D pictures of unique objects such as viruses, organelles or cells. This is a protocol for macromolecular structure determination from tomographic data using subtomogram averaging in the RELION software.
Ramani et al. describe a protocol for in situ DNase Hi-C as an alternative to traditional Hi-C methods that use restriction enzymes. The use of DNase I for chromatin digestion circumvents the resolution limit imposed when relying on genomic restriction sites.
G&T-seq enables sequencing of DNA and mRNA from the same single cell, allowing the effects of genomic variation on transcription to be studied. It is compatible with any WGA method and so can be tailored to specific applications.
This protocol describes a simple pipeline for imaging FRET biosensor probes with two-photon laser scanning microscopy (TPLSM). Validated FRET standards are described that can be used with any TPLSM setup to ensure sensitive detection of FRET ratios.
This protocol from Nguyen et al. describes the use of the plant cyclase butelase 1 for the efficient cyclization and ligation of peptides and proteins. After extraction from Clitoria ternatea, the protocol describes reactions for cyclization, ligation and synthesis of protein thioesters.