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A Deep-sequencing-assisted, Spontaneous Suppressor Screen in the Fission Yeast Schizosaccharomyces pombe
JoVE Journal
Genetik
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JoVE Journal Genetik
A Deep-sequencing-assisted, Spontaneous Suppressor Screen in the Fission Yeast Schizosaccharomyces pombe

A Deep-sequencing-assisted, Spontaneous Suppressor Screen in the Fission Yeast Schizosaccharomyces pombe

DOI:
10.3791/59133-v

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07:55 min

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March 07, 2019

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, ,
1Department of Biology,Wake Forest University, 2Center for Molecular Communication and Signaling,Wake Forest University

Kapitel

  • 00:04Titel
  • 00:32Plate Reader Assay
  • 03:13Selection of Suppressor Colonies and Confirmation of Phenotype
  • 05:27Results: Slow-growing Mutants Show Phenotypic Recovery and Genetic Confirmation of Identified Suppressor Mutations Verifies the Heritability of the Recovery Phenotype
  • 06:56Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

We present a simple suppressor screen protocol in fission yeast. This method is efficient, mutagen-free, and  selective  for  mutations that  often  occur at   a  single  genomic  locus.  The protocol is suitable for isolating suppressors that alleviate growth defects in liquid culture that are caused by a mutation or a drug.

Tags

Keywords: Fission YeastSchizosaccharomyces PombeSuppressor ScreenDeep SequencingSpontaneous MutationGrowth DefectOptical DensityPlate ReaderAutomated DilutionKinetic Growth Curve

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