Liquid food consumption

The researchers will need to prepare S media with OP50 culture, 500uM FUDR, 1X Antibiotic-Antimycotic, and 96 well plates. Additionally, it is necessary to prepare L4 synchronized animals. The researchers need to use a multi-channel pipette to rapidly mix solution before using a multimode plate reader (e.g. BioTek Synergy H4 multimode plate reader). 

1) Prepare S media containing 500 uM FUDR and 1 x Antibiotic-Antimycotic (ThermoFisher 15240062).  Add E. coli OP50 to the prepared S media to density OD600 of 1.0,  Mix the OP50 completely before aliquoting 150uL to each well of the 96 well plates. The researcher may use 16 - 24 wells for each strain.

2) Have the synchronized animals ready. Use S media wash off the animals off the plate into a 1.5mL tube. Wait for 2 minutes. Remove the supernatant. Wash animals with S media containing OP50 (OD600 = 1), 500 uM FUDR and 1 x Antibiotic-Antimycotic for 3 times.  Transfer 1mL animal suspension on a 3 cm petri dish. Transfer 20 animals to each well on the 96 well plate using a pipette (P20 should be good to use). Use a dissecting microscope to check the animal number in the well of 96 well plate.

3) Using  multi-channel pipette to completely mix the well before using the multimode plate reader and get the Day 1 OD600 values.

4) seal the plate using a plastic film. Place the 96 well plate in 20C incubator.

5) read Day2-Day5 OD600 every 24 hours by a multimode plate reader. Before reading, remove the film and completely mix the wells by using multi-channel pipette.