Culturing and growing worms

Model Organism: Worms Methods & Reagents
1

Hello,

Does anyone have experience with microwaving NMG after it has already been autoclaved and solidifies to pour plates? As in make some NGM today and then use it over the next few days to pour plates by microwaving it to liquify instead of making fresh NGM and autoclaving each day. Does this work? If yes, can you add all of the additives to it from the very beginning or just the three ingredients that go into it before you autoclave and then add the additives each time after you microwave?

On another note, for bleaching worms - dI water vs. M9 vs S-basal what are the pros and cons for washing down the worms during bleaching in each of these? And which is the better solution to hatch eggs into L1s prior to pipetting them to a seeded plate and why? End goal is the adult population that will hatch from these L1s to use for experiments.

Thank you!

2

After bleaching (and all liquid handling of worms including hatching), we use 50 mM NaCl since we know that M9 changes the expression of the genes we are most interested in:

http://wbg.wormbook.org/2012/06/25/m9-and-nlp-29-standard-buffer-can-induce-antimicrobial-peptide-gene-expression/

3

I would recommend heating the premade media in a 65C water bath rather than microwaving it.
Although I have not done this, it should permit you to add the heat-sensitive components before storage.
It should be relatively easy to test whether the plates are good by seeing if they “work.”
When you microwave a solid like NGM in a standard microwave, the media heats unevenly.
The little pockets of melted media can get too hot for any additives.
My thoughts, Janet

4

Thank you both!