I have a very basic question. one of my assay is not working and i am beginning to question my fundamentals. N2 worms are grown on NGM plate fed with OP50 at 20 C. My question is do the worms start laying eggs 72 hours post-bleaching?
I don’t see why they shouldn’t have laid eggs by then. Typically we look at animals staged from the late-L4 stage. Typically ~20 eggs have been laid per adult animal at 18 hours post-L4 on OP50 at 20°C. Most strong Egl phenotypes are obvious at that point, although when we quantitate these defects we wait until 30, or sometimes 40 hours. After 30 hours, Egl animals may have hatched progeny, confounding results.
-Kevin.
At 20C, the N2 worms reach the egg-laying adult stage 72h post-hatching. If you count time from bleaching, you have to add 8-12h for
embryo/morphogenesis. In my hands, 3 days after I remove the parent from the plate (equivalent of bleaching), I have a mixt population of
young adult and egg laying adult.
Hi Visygen, I noticed that you synchronized your worms by bleaching. Normally, most embryo (eggs) from bleaching are at earlier stage than laid egg. For laid eggs, it takes about 3.5 days for them to reaching gravid adult at 20 deg. C. So maybe you need one more day in your assay to ensure most of your worm are gravid.
@magicper, Well the problem with me is actually that worms are in an advanced age than they are supposed to be.
I have described the exact nature of my problem in a separate post i have put up now under the titel “Problem with worm age and behavior”