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