Yeast Spotting Assays
Yeast Spotting Assays - Related Videos
09:06
Growth Assays to Assess Polyglutamine Toxicity in Yeast
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Cited by 3 •
2012
This manuscript describes three complementary protocols for assessing the toxicity of polyglutamine (polyQ)-expansion proteins in the yeast Saccharomyces cerevisiae. These protocols can easily be modified to monitor the toxicity of other misfolded proteins in yeast.
05:20
Yeast Two-Hybrid Assay to Determine Protein Self-Association in Yeast Cells
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2025
This video demonstrates the yeast two-hybrid assay to detect specific protein self-interactions. In this assay, the protein of interest is tagged to two domains of a transcription factor that encodes the β-galactosidase enzyme. When the two proteins interact, this causes transcription and translation to form the enzyme, which is then detected using a biochemical assay.
03:08
Light Spot Assay: A Method to Investigate Drosophila Phototactic Behavior
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2023
Drosophila larvae are light-sensitive and respond to changes in surrounding light. This video describes an assay that tests their ability to avoid light, called the light spot assay. The featured protocol demonstrates how to set up the assay with a blue light LED and shows how to record the animal's behavior in response to the light encounter.
10:27
A Fluorescence Microscopy Assay for Monitoring Mitophagy in the Yeast Saccharomyces cerevisiae
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Cited by 26 •
2011
A robust approach to monitor the delivery of organelles to the acidic lumen of the yeast vacuole for degradation and recycling is described. The method relies on the specific labeling of target organelles with a genetically encoded dual-emission fluorescence pH-biosensor, and visualization of individual cells using fluorescence microscopy.
06:39
Real-Time Void Spot Assay
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Cited by 14 •
2023
This article describes a new method to study mouse voiding behavior by incorporating video monitoring in the conventional void spot assay. This approach provides temporal, spatial, and volumetric information on the voiding events and details of mouse behavior during the light and dark phases of the day.
FAQs
The dilution series shows whether a growth difference persists as the number of plated cells decreases. A strain that forms stronger spots across several dilution levels demonstrates a different growth or survival pattern from a strain with faint or absent spots. Examining the full series therefore provides more informative comparisons than judging a single spot concentration.
Spot intensity provides a visual measure of colony formation under the tested condition. Stronger spots indicate more apparent growth or survival, whereas weaker spots indicate reduced performance relative to the comparison. Interpreting intensity across multiple dilution levels helps reveal the extent of a strain’s response rather than relying on one visual observation.
Applying defined chemical or environmental challenges can expose differences in how yeast strains maintain growth or survival. Temperature, osmotic stress, nutrient limitation, and compound sensitivity may produce distinct spot patterns between strains. Comparing these responses helps connect particular genetic backgrounds with cellular physiology and can provide evidence about molecular mechanisms affecting adaptation or viability.
A typical workflow begins with yeast cultures, followed by preparation of serial dilutions. Each dilution is spotted onto solid agar containing the defined experimental condition, and the plates are incubated. Researchers then compare colony formation and spot intensity across the dilution series, strains, or conditions to identify changes in growth or survival.
The essential materials are yeast cultures, serial dilution preparations, solid agar, and plates that impose the selected defined condition. The assay can be adapted by changing the strain, chemical exposure, temperature, osmotic environment, or nutrient availability. Keeping the comparison conditions defined allows differences in spot patterns to be associated with the tested variable.
Researchers can use the assay to compare yeast strains, investigate gene function, or measure sensitivity to compounds and environmental stresses. Its visual readout supports studies of cellular physiology and molecular mechanisms while requiring relatively simple, inexpensive materials. The approach is especially useful when many strain or condition comparisons must be evaluated through their growth patterns.