Lidstrom:EMS Mutagenesis

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(New page: Back to Protocols == Overview == * EMS is a liquid mutagen ** '''Work in the fume hood and use special precautions!''' * It tends to produce a certain type of mutat...)
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Revision as of 09:07, 26 June 2014

Back to Protocols

Contents

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.


Sample Protocol

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
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