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Protocols and programs for high-throughput growth and aging phenotyping in yeast.

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Summary

This study enhances yeast phenotyping by adapting 96-well plate growth assays to 384-well plates, improving throughput for growth rate and Chronological Life Span (CLS) determination. New software, GATHODE and CATHODE, aids in analyzing these high-throughput yeast experiments.

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Area of Science:

  • Microbiology
  • Systems Biology
  • Biotechnology

Background:

  • Standard yeast phenotyping relies on growth rate determination in various conditions.
  • Current high-throughput methods use 96-well plates for growth curve analysis.
  • Chronological Life Span (CLS) determination is typically a laborious Colony Forming Unit counting method.

Purpose of the Study:

  • To adapt and validate a 384-well plate microcultivation method for higher-throughput yeast growth assays.
  • To develop and validate a higher-throughput assay for yeast Chronological Life Span (CLS).
  • To create software tools for efficient analysis of large datasets from these assays.

Main Methods:

  • Adaptation of 96-well plate growth curve assays to 384-well plates.
  • Validation of growth parameters (lag time, doubling time, biomass yield) against 96-well plates and batch cultures.
  • Development of a 384-well plate assay for yeast Chronological Life Span (CLS).
  • Creation of GATHODE and CATHODE software for semi-automatic data analysis.

Main Results:

  • Growth parameters from 384-well plates showed strong correlation with 96-well plates and batch cultures.
  • The 384-well plate method demonstrated consistent results across different yeast species.
  • A validated higher-throughput CLS assay was established.
  • GATHODE and CATHODE software facilitate efficient analysis of growth and CLS data.

Conclusions:

  • The 384-well plate microcultivation method significantly increases throughput for yeast phenotypic screens.
  • The developed assays and software enhance the time- and cost-efficiency of yeast systems genetics experiments.
  • This approach is applicable to various yeast species and screening applications.