Lidstrom:EMS Mutagenesis

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Overview

• EMS is a liquid mutagen
  • Work in the fume hood and use special precautions!
• It tends to produce a certain type of mutations

Tips

• Use enough cells that you can see the biomass pellet after spinning down the culture to remove the EMS solution.
• Use clear eppendorf tubes to hold the cells in so you can see the biomass pellet.
  • One could use opaque tubes to reduce EMS's reaction with light, but the protection from light probably isn't worth the convenience of

Health & Safety

• EMS is a liquid mutagen
  • Work in the fume hood and use special precautions!
• EMS can be slowly inactivated by mixing with a sodium thiosulfate + NaOH mixture.
  • Recipes found online vary a bit, but they are about _____:
• Leave all plastic tips and gloves used for the experiment in the fume hood a day or so after use. Double bag them and discard them in the normal trash stream after that.

Open Questions

• Should you spin down the cells to remove the culture supernatant they were growing in?
  • Yes, if:
    • it has food you don't want them to have access to after mutagenesis
    • you are concerned that EMS could react with compounds in the media
    • you aren't in a rush and want to lean toward a more consistent but time consuming workflow.
  • It is probably fine to skip this step most of the time though.

Misc Notes

• The fact