layout title date default Prototols 2020-12-01 Molecular cloning Gibson assembly Golden gate assembly Freezing and streaking bacteria Miniprep (regular) Miraprep (enhanced miniprep) Gel purification PCR amplification Commonly used E. coli genotypes Cell and organ culture Useful cell numbers in various culture formats Routine cell culture Freezing and thawing cells 3D spheroid cell culture Embryonic salivary gland culture Fluorescence-activated cell sorting (FACS) Cell cloning by limited dilution Cell line transgenesis Lentivirus packaging Lentivirus transduction PiggyBac-mediated transgenesis Transcription modulation Inducible CRISPRi and CRISPRa Guide RNA design for CRISPRi and CRISPRa Gene knockdown with CasRx Guide RNA design for CasRx Genome engineering Guide RNA design Gene knockout in cell lines Small fragment knockin in cell lines Large fragment knockin in cell lines Gene knockout in C. elegans Small fragment knockin in C. elegans Large fragment knockin in C. elegans Gene knockout in mice Small fragment knockin in mice Large fragment knockin in mice Immunostaining Whole-mount immunostaining for mouse embryonic organs or 3D spheroid cultures Single-molecule RNA fluorescence in situ hybridization (smFISH) smFISH for tissue culture cells smFISH for whole-mount mouse embryonic organs Microscopy Imaging of immunostained cells and tissues Live-cell imaging Live-organ imaging Microtubule gliding assay Oxygen scavenger Fluorescence dye labeling Fluorescence dye labeling of proteins Fluorescence dye labeling of DNA oligos Bulk and single-cell RNA-seq RNA extraction using Trizol Tissue dissociation into single cells Protein biochemistry Buffering reagents His-tag protein purification MBP-tag protein purification GST-tag protein purification Gel filtration Sucrose gradient Siegel-Monty oligomerization analysis Microtubule polymerization Microtubule co-sedimentation Microtubule co-sedimentation from worm exacts