aliquoting worm eggs

Model Organism: Worms Methods & Reagents
1

Does anyone have suggestions for how to reliably aliquot approximately the same number of eggs to multiple plates following bleaching of adults? I’m trying to compare multiple RNAi clones simultaneously and want to get about the same number of growing worms per RNAi plate.

Even with frequent mixing, the eggs in M9 distribute pretty non-homogeneously and I end up getting a lot of variation in eggs per plate after aliquoting. I’m wondering if anyone has resuspended eggs in something more viscous to prevent settling?

Any advice would be greatly appreciated.

2

You’ll get much more reproducible results dispensing L1s instead of embryos. Bleach the gravid adults, hatch overnight in the absence of food, and resuspend in water or M9. Determine the density of worms (typically by counting the number in a 10 µl drop), adjust if necessary, and dispense by repeating pipet. Shake your worm suspension each time before loading the pipet. Variation is typically <25% between plates.

Good luck!