culture media

Leon Avery leon at eatworms.swmed.edu
Thu Nov 9 08:26:44 EST 1995


In article <47s29m$rn2 at dingo.cc.uq.oz.au>, L.Eden at tpp.uq.edu.au (Lois Eden) writes:
|> 
|> 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

Agar contains enough sterols for survival, but for optimum health,
especially at low temperature, you need to add some.  (Cholesteroal
doesn't cost much, and you don't need much, and it's stable to
autoclaving, so why not add it, anyway?)  This may be the reason for
your restricted temperature range--most C elegans people have no
trouble growing the worms at temperatures between 12 and 25C.

Chunking is a useful way of transferring worms, but over the long term
you're likely to contaminate your stock with airborn yeast or
bacteria.  We mostly use platinum picks (not expensive--the amount of
platinum is tiny) to transfer single animals.

Media recipes and other methods can be found in several places: the
Ambros Lab Comprehensive Protocol server
http://www.dartmouth.edu/artsci/bio/ambros/protocols.html, the Cold
Spring Harbor book "The Nematode Caenorhabditis elegans", edited by WB
Wood et al, and the recently published "Caenorhabditis elegans: Modern
Biological Analysis of an Organism", Epstein and Shakes editors, from
Academic Press.  

It might be a good idea to put some of this information in the FAQ.  I
guess I should write up a more complete answer (unless someone else
feels like it).

-- 
Leon Avery					   (214) 648-2420 (office)
Department of Biochemistry			            -2768 (lab)
University of Texas Southwestern Medical Center             -8856 (fax)
5323 Harry Hines Blvd				   leon at eatworms.swmed.edu
Dallas, TX  75235-9038			  http://eatworms.swmed.edu/~leon/



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