Modified dauer purification protocol

From Wikionchus

1. wash off approx. 10 big plates (10cm diameter) full of dauer produced using the Wet Plate Protocol for the production of "dauer cultures" with M9/Triton (0.2%) and transfer them into one 50 ml tube

2. spin at 1300g for 2 minutes

3. remove the supernatant, resuspend the pellet with 40 ml Ficoll (20% in M9/Triton)

4. wait for 5 minutes

5. distribute the Ficoll solution into 2 50 ml Falcon tubes and fill them up with M9/Triton

6. spin at 1300g for 2 minutes

7. remove the supernatant, resuspend the pellet with 40 ml Ficoll (20% in M9/Triton)

8. spin at 1300g for 2 minutes

9. remove the supernatant, resuspend the pellet with 20 ml Ficoll (20% in M9/Triton) and add 30 ml Sucrose (50% in M9/Triton)

10. spin at 1300g for 10 minutes

11. transfer supernatant (containing dauers) into two 50 ml Falcon tubes and resuspend with 50 ml M9/Triton (dauers are not happy in sucrose). Dispose pellet with eggs and debris

12. spin at 1300g for 10 minutes

13. remove supernatant and combine both small pellets (full of pure dauers) with 30 ml M9/Triton