HI, I’d like to post some steps of my experiment to share and to get advice.
1 For life span analysis, the plates should be filled at least for 1/2 of the capacity. I use Corning’s 60mm plates for my experiment, and usually it needs 6ml NGM.
2 To prevent the starvation, 100ul OP50 is gentlely added to the plates in day3, day6, day12, day16. Slowly and gentlely swirl the plates to let the fluid spread well -distributed in the plates.
3 In day8 and day18 transfer the worms in the new plates. Based on my experience, picking is not a good method for the worms flappy and becoming weakness. I use small agar cubes(~4mm*4mm) with thick bacteria lawn cut from an old NGM plate which is a little hard to stick the worms as a pick. It usually transfer 2-3 worms a time and it works quickly. When put the cube with sticking worms to the new plates, put it down on the non bacteria area to avoid disturb the lawn.
Thank you
I have searched the stardard life span test protocal for a long time, but I have not found it. Is it because the test is too simple to put it on? I have no idea because our lab even do not know the worms and I’m the first guy culturing worms.
when the adults lay eggs, the worms are transferred to fresh plates daily, to avoid mixing the generations.(Couillault, C. and Ewbank, J. J.,2002)
I think you will find the answer from the papers about the microbes and infection.
Thank you, anne!
For my life span analysis, all plates are added the FUdR to avoid progeny.First, I used 100ug/ml for a lot of paper used this dose. And then I found 12.5ug/ml also worked good(Mohan Viswanathan etal,Developmental Cell, Vol. 9, 605–615, November, 2005).