Hello all.
I was wondering if anyone could provide or point me in the right dirrection to get a reliable and reproducible hydrogen peroxide resistance assay.
Preferably one with details on how to determin weather a worm is dead or not when suspended in a liquid.
Cheers
Shaun
Hi,
Pamela Larsen used a peroxide assay in a paper from 1993, perhaps you have this already;
http://www.pnas.org/content/90/19/8905.full.pdf
She scored dead/alive worms in 96 well plates by movement (e.g. do they move if the plate is tapped) and for non-swimmers, by looking for pharyngeal movement). She had av. 35 worms per well.
Then there’s always dyes such as SYTOX (Invitrogen) which is taken up by dead worms, but I’m not sure whether that would work in the presence of peroxide…I think the visual methods are ok anyway.
Regards
Steve
I came up with an assay to measure the resistance to the effect of hydrogen peroxide treatment of starved L1s on subsequent development. It is easy to do and quantify. daf-16 mutants are very sensitive and daf-2 or age-1 mutants are resistant. The reference is http://www.biomedcentral.com/1741-7007/4/1. If you have any questions, please let me know.
David