1. Melt freezing solution in microwave (4 minutes at 70% power works)
2. Place in 50°C water bath until ready to use.
3. Wash worms with M9 into 15ml conical tube
4. Spin for about 20 seconds to pellet worms
5. Aspirate off to 2ml
6. Put tube on ice for 20-30 minutes
7. Add 2ml freezing solution to worm tube
8. QUICKLY pour up to line in freezing vials
9. Put vials at -80°C overnight
10. Test thaw a pea-sized piece on a small plate. This may take several days.
11. If test thaw works, keep one vial in the –80°C freezer, and put the other in liquid nitrogen.
12. Be sure to note in database when the strain was frozen, if the test thaw was successful, and which box (in both -80°C and liquid N2) the vial is located.
* I always keep a plate of worms just in case the test thaw fails.