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Cell Biology and Research Processes

A compilation of diagrams and flowcharts related to cell biology, including setup MiniStats with various tubing sets, budding polarity assay, and mitochondrial biosensor imaging in budding yeast.

Graphical Abstract describing growth, imaging, quantification of mitochondrial biosensor expressing cell.

Graphical Abstract describing growth, imaging, quantification of mitochondrial biosensor expressing cell.

Generation of mtHyPer7-expressing cells by CRISPR. The Cas9 and sgRNA-containing plasmid (YN2_1_LT58_X2site) and PCR-amplified mtHyPer7-containing biosensor construct are introduced into budding yeast cells by lithium acetate transformation. The gene-free insertion site on chromosome X (X2) is recognized and cut by Cas9 protein with the sgRNA, and the biosensor is integrated into the genome by homologous recombination. After identification of the successful transformants by microscopic screening, colony PCR, and sequencing, the Cas9 plasmid is removed (cured) by growth in non-selective media. Abbreviations: sgRNA = single guide RNA; TEF = transcription enhancer factor.

Generation of mtHyPer7-expressing cells by CRISPR. The Cas9 and sgRNA-containing plasmid (YN2_1_LT58_X2site) and PCR-amplified mtHyPer7-containing biosensor construct are introduced into budding yeast cells by lithium acetate transformation. The gene-free insertion site on chromosome X (X2) is recognized and cut by Cas9 protein with the sgRNA, and the biosensor is integrated into the genome by homologous recombination. After identification of the successful transformants by microscopic screening, colony PCR, and sequencing, the Cas9 plasmid is removed (cured) by growth in non-selective media. Abbreviations: sgRNA = single guide RNA; TEF = transcription enhancer factor.

Tubing assembly in mCAD.

Tubing assembly in mCAD.

Preparation of mCAD components for autoclaving

Preparation of mCAD components for autoclaving

mCAD components

mCAD components

Schematic of the fully assembled mCAD

Schematic of the fully assembled mCAD

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