culture media

[Lois Eden]

In article <wfrice00-0711951746400001 at dialin3.slip.uky.edu>, 
wfrice00 at pop.uky.eduÊ says...
>
>     I am a first year graduate student beginning studies in hatching 
of
>C. elegans eggs.  Does anyone have a favorite recipe for a good 
culture
>media?  We have found several different nutrient combinations in 
texts,
>but I would like to get some input regarding which work the best.  
Also,
>does a standard nutrient agar mix contain enough cholesterol to 
maintain
>the worms, or must additional sterols be added to the mix?  Any reply
>would be greatly appreciated.
>
>Wayne Rice 
>University of Kentucky, Lexington

I have been maintaining a culture since Nov 1990.  I use 1/4 strength 
nutrient broth (Difco) with an agar content of 2% (so the nematodes 
cannot burrow into the media).  Plates have E.coli lawns on them before 
transferring nematodes onto them.  They have to be maintained at 20-23 
degrees or they die out (this may vary with the strain of C. elegans 
you use).  You should transfer them about once a week to keep high 
numbers up.  Transfer time can vary depending on how many nematodes you 
transfer.  I transfer by cutting an agar square out of one plate and 
putting it onto the next.  I have no idea about sterols etc in standard 
nutrient mix but my nematodes are alive and thriving after 5 years, so 
it must be OK.

Lois Eden
University of Queensland