worm washing

hello!
Worm papers often refer to washing of worms. Is there a standard procedure for this, ie what rpm? for how long? thanks for any advice!

We usually wash at 2000rpm (320xg) for 2 mins and the washing is repeated 3 times.

I agree, spin at 2000 RPM for 4-5 minutes. You can used M9 with 0.1% Triton-X initially to clean up any bacteria, then wash a few times with just M9 or dH20.

Hello, I’m infecting worms with different species of Candida and then mount a microplate assay with 20% BHI, 80% M9 and kanamycin. The problem presented to me is that Candida proliferates, resulting in turbidity (although I washed the worms as recommended and I allowed it to crawl on BHI agar).
I thought that perhaps the absence of cholesterol in ethanol (by an oversight) :-[ could influence. :o
If someone could guide me I would make a great favor.
Thanks. :slight_smile:

Do washing kill the worms?
Or those worms can be further used for assays?

Hi, I’m sorry for not responding earlier.

The protocol is as described Breger et al, Briefly:

Approximately 100 worms (L4 stage) were dispensed onto a lawn of Candida and allowed to feed for 2 h. The worms were remove off the plates with M9 buffer and wash at 1200 rpm for 2 mins (the washing is repeated 3 times). The pellet of worms was allowed to crawl on unseeded BHI plates to remove yeast cells from their cuticles… but this procedure was not successful because when I pass the worms to a liquid medium in a microplate (79% M9 buffer, 20% BHI, 10 ug/ml cholesterol in ethanol, and 90 ug/ml kanamycin), Candida also grows and the medium show turbidity, and at 48 hours I can not see anything

how long should I allow the worms to crawl, so that your cuticles are well clean?

I am sorry for so poorly writing in English and thank you very much.