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High Throughput Yeast Strain Phenotyping with Droplet-Based RNA Sequencing
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Published on: May 21, 2020

Genotyping 1000 yeast strains by next-generation sequencing.

Stefan Wilkening1, Manu M Tekkedil, Gen Lin

  • 1Genome Biology Unit, European Molecular Biology Laboratory, Meyerhofstr. 1, 69117, Heidelberg, Germany.

BMC Genomics
|February 12, 2013
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Summary

A new, cost-effective library preparation method enables high-throughput whole-genome sequencing of yeast. This approach significantly reduces costs and time, facilitating large-scale studies of recombination and aneuploidy.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Yeast Genetics

Background:

  • Next-generation sequencing (NGS) throughput has surged, but library preparation remains a bottleneck.
  • Current methods are costly and time-consuming for large sample numbers.
  • A need exists for economical, high-throughput library preparation for NGS.

Purpose of the Study:

  • To develop and validate an economical, high-throughput library preparation method for Illumina sequencing.
  • To enable large-scale genomic analysis of yeast populations.
  • To investigate meiotic recombination and aneuploidy in yeast.

Main Methods:

  • Developed a 96-well based DNA isolation protocol for yeast.
  • Implemented a low-cost DNA shearing alternative.
  • Utilized heat inactivation for adapter ligation, omitting bead cleanups.
  • Applied the method to prepare up to 384 whole-genome libraries weekly.

Main Results:

  • Achieved library preparation for under 15 euros per sample.
  • Successfully sequenced over 1000 yeast genomes at ~30x coverage.
  • Generated a high-resolution meiotic recombination map from 768 yeast segregants.
  • Detected 3.6% aneuploidy and identified chromosome missegregation patterns.

Conclusions:

  • Presented a cost-effective, high-throughput workflow for yeast genome sequencing.
  • The method facilitates large-scale studies of recombination and aneuploidy.
  • Provides a foundation for future research on linkage and chromosomal aberrations in yeast and other eukaryotes.