Dead Worms on EMS...

Model Organism: Worms Methods & Reagents
1

Hi all,

Thanks for your help in advance. I just performed an EMS mutagenesis using several hundreds Po animals at an EMS concentration of 50mM (4mls of M9 gently rocking for 4hrs). At the end something like 95% of my worms were dead. Is that normal? They were all sticks…very discouraging.

Are these results typical? I am mutagenizing a mutant, but its not that sickly of a strain, so its confusing. I should mutagenize N2 as a control, but thought I’d ask the general community about this first.

Cheers…

2

I’ve always used a wheel rather than gentle rocking, but that’s not at all normal. I’m not sure offhand of the EMS concentration I used (it’s 20ul EMS mixed well in M9 before you add the worms in M9 to a total volume of 4ml, but I don’t know the molarity), but even moderately sick elegans strains fared well; certainly, few if any were dead afterwards. I suppose it could be the rocking, or something in the EMS, but I’d bet it’s the strain. In any case, you could know in about 4 hours.