High-throughput screening of multiple Caenorhabditis elegans strains

Prepare 60 mm NGM plates for worm maintenance. Dry plates for 1 hour under flow hood. If using 40 strains per week, you need at least 120 seeded plates per week (chunking Monday, Wednesday and Friday).

Always use the same agar, peptone and LB throughout the project (do not change reagents mid project).

Prepare overnight cultures of OP50 E. coli by picking one colony into a 50 mL Falcon tube containing 30 mL LB Miller. Incubate overnight at 37ºC with 200 rpm shaking.

Seed plates with 200 μL liquid culture. Keep plates in cold room until ready to use.

Prepare 24 well plates with peptone free NGM by dispensing 600 μL per well using Integra VIAFILL dispenser. Dry plates for 1 hour under hood. Prepare enough 24 well plates for 2 weeks and keep them in cold room until use. Seed plates with 15 μL OP50 per well the day before experiment.

Get strains out of freezer and grow them for at least a week without starving. I chunk worms every Monday, Wednesday and Friday. Make a very small chunk on Friday so that they are not starving the following Monday.

Have worms on 60 mm plates with plenty of food and eggs. 24 well plates need to be seeded with 15 μL OP50 the day before and left on bench. Pick 5 to 20 eggs per well. Use 2 rows per strain and save the plate maps to make the metadata. Keep plates at 20ºC for 3 days.

• If there are not enough eggs, pick an adult worm and allow it to lay eggs on wells. Remove the worm.

• Pick eggs Monday, Tuesday and Friday for imaging Thursday, Friday and Monday.

Allow eggs to hatch and grow on plates for 3 days. Worms should be day 1 adults.

Acclimate worms on imaging room for at least 30 min. Start imaging 24 well plates.