I’m hoping to get some very basic advice on how to use FUDR: do you make a stock solution and dilute into the plate agar when you pour the plates, or add it as a solution to already cooled plates and let it soak in? I’m looking for very basic details such as stock solution concentrations, what you make solutions up in, final concentration that is most effective, etc.
Also, if you place worms as L4s, will that allow enough time for the FUDR to inhibit all progeny production, or would you suggest placing, e.g., L1s down onto the FUDR plates?
I have used FUDR in C elegans life span experiments for a long time. I make a stock solution 10 g/L in DD H2O, and add it as a solution to hot agar(65-80 degree centigrade). The final concentration is 25mg/L(8mM). I place worms as L4s-young adults. It works well. I bought the reagents from sigma, CAS46875.
However, in my new experiments, FUDRdid not work even in 100mg/L. The fudr was also bought from sigma, CAS F0503.
Did somebody meet such a problem?
Is the FUDR sensitive to heat? When I make RNAi plates and selection plates I always wait until the agar is cool enough to keep my hand on the glassware without it being painful. Although I am forced to wait longer than I would like, I never have any problems. It would be unfortunate if it is problem this the chemical itself.