Synchronisation of nematodes

1. wash mixed stage worm plates heavily with M9 into 50ml Falcon tubes

2. centrifuge for 3 minutes at 1300g

3. discard supernatant

4. add 15 ml 1:1:3 NaOH(1N), Chlorix and aqua dest.

5. leave for 10 minutes, vortexing every 2 minutes for 10 seconds

6. centrifuge for 3 minutes at 1300g

7. discard supernatant

8. add 25 ml NaCl 0,1M, shake, then leave for 5 minutes

9. centrifuge for 3 minutes at 1300g

10. discard supernatant

11. add 20 ml M9, shake gently

12. centrifuge for 3 minutes at 1300g

13. discard supernatant

14. add 20 ml M9

finally put on shaker for 1-2 days