Life span assay

Please I need advice or experienced suggestions from forum members on the addition of 5-fluorodeoxyuridine (5-FUD) into my NGM plates for carrying out the lifespan experiment. I wish to know what volume of the 5-FUDR would be enough for preventing hatching of progeny of the worms. For example if I make a 750 ml volume of NGM? Also, I’d like to know if the 5-FUDR is better added after allowing the media to cool down from autoclaving at the same time am adding cholesterol, Calcium and Magnessium or it can be added to the NGM plates having poured and allowed them to solidify overnight? Thank you.

I’m sure that there are a number of sources for this info, but I found the this JOVE methods paper from George Sutphin and Matt Kaeberlein to be quite helpful and should address many of your questions.

But note the caveats mentioned in the following post:
Best of luck with the lifespan assays,

Thank you all for the answers. They have been helpful indeed.