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EMS Mutagenesis

  1. Add 4 ml M9 per plate to 1 or more plates of worms with many L4's.
  2. Suck up worm suspension in a pasteur pipet and place in a 15 ml falcon tube.
  3. Wash the worms by spinning 30s at 1200 rpm, remove supernatant and bring volume to 3 ml.
  4. Add 1 ml of a solution of 20µl EMS / 1ml M9. (de-glove after opening and closing EMS bottle).
  5. Mix, close tube and wrap with parafilm.
  6. Rotate tube at 15°C or 20°C for 6 hrs.
  7. Wash the worms 3 times with 4 ml of M9 buffer. Use disposable plastic pasteur pipettes.
  8. After last wash, pour of supernatant and suspend worms in remaining liquid.
  9. Add worm suspension to plate and allow liquid to dry into plate.
  10. After liquid has dried into plate, pick L4's.



  • Watch tge temperature if using temperature sensitive mutants.



  • Put EMS containing liquids in saturated NaOH. After 1 day dispose like normal waste.
  • Put rest (gloves, tips, plastic pipettes) in solution of 400 g sodium thiosulfate in 4 L water. After 4 days dispose like normal waste.