calorie restriction

Hi,

Does anyone have a protocol to do calorie restriction? I am trying to test whether CR can affect some of my mutants. Thanks very much.

Shouhong

There are a number of different protocols, and they have some very distinct (and annoying) differences. Basically, you can either use ordinary NGM plates or liquid culture (with a few significant variations). With plates, you’re closer to ordinary growth/assay conditions, so that’s where you might to start. Basically you seed plates with dilutions of live OP50, and transfer worms to fresh plates every day or two so the OP50 doesn’t have a chance to grow up. You can also use dead OP50, so you don’t have to transfer the worms as often (though they’re still transferred in the published experiments I’ve seen, for consistency with the live OP50 CR). Look up the papers from Anne Brunet’s lab – they developed the solid dietary restriction protocol, and they outline it pretty thoroughly in the methods. There’s also another paper from her lab that compares the various other methods – you can probably find the liquid CR methods by hunting down its references.

Thanks very much.

shouhong