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Yeast Protocol pp 121-127 | Cite as

Mutagenesis

  • Leslie Barbour
  • Michelle Hanna
  • Wei Xiao
Part of the Methods in Molecular Biology book series (MIMB, volume 313)

Abstract

To identify new genes in an organism, a genetic approach can be used to screen for mutations that display a particular phenotype. Genotoxic agents, such as ultraviolet (UV) light, ionizing radiation, or chemicals can be used to randomly induce DNA lesions in the genome. Most efficient mutagenesis occurs when a mutagen confers a high frequency of mutations with low lethality, in the range of 10 to 50% survival. These mutations can be in the form of frameshifts, deletions, or rearrangements. To initiate a mutagenesis, a fresh subculture of cells grown into log phase is collected, washed, and resuspended in potassium phosphate buffer. The mutagen is added to the culture for a predetermined time, deactivated, and washed from the cells. The cells are allowed to recover from the treatment by incubating in liquid or on solid medium. Mutants can be isolated by screening individual colonies or by using direct selection of cells from the mutagenized cell population.

Keywords

Yeast Strain Mutation Frequency Fume Hood Forward Mutation Mutagenesis Protocol 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Humana Press Inc., Totowa, NJ 2006

Authors and Affiliations

  • Leslie Barbour
    • 1
  • Michelle Hanna
    • 2
  • Wei Xiao
    • 2
  1. 1.Institute of BiotechnologyUniversity of SaskatchewanSaskatoonCanada
  2. 2.Department of Microbiology and ImmunologyUniversity of SaskatchewanSaskatoonCanada

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