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.
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.
- Yes, if:
Misc Notes
- The fact