100,000 is not a large number of worms. There are about that many
worms on a single starved 6 cm plate (but almost all are L1s). If
you need adults, you could easily get that many from 20 10 cm
enriched plates. By growing them in liquid (which is a bit more of a
chore, but still routine) you can get millions.
"Totally free" of bacteria is pretty demanding -- what are you
planning that would be screwed up by 1 bacterium mixed in with
100,000 worms? You can get eggs free of bacteria by treatment with
basic hypochlorite. You can get newly hatched L1s free of bacteria
by letting the eggs hatch in sterile buffer. In principle you could
grow those worms in axenic medium -- such media exist, but they are
really crummy. You can also grow up a lot of worms in bacteria, then
wash them in sterile buffer as many times as you like, to approximate
a population free of bugs. You could use bacteria that were
sensitive to streptomycin and kanamycin, wash away almost all the
bugs, then do whatever experiment you have in mind in buffers
containing strep and kan, to kill the few remaining ones.
--
Leon Avery (214) 648-4931 (voice)
Department of Molecular Biology -1488 (fax)
University of Texas Southwestern Medical Center
6000 Harry Hines Blvd leon at eatworms.swmed.edu
Dallas, TX 75390-9148 http://eatworms.swmed.edu/~leon/
>Hi,
>>I've scanned previous posts to this group and lots of websites pretty
>thoroughly but can't find answers to a couple of questions that I
>have, so I hope someone can either help me or point me to a suitable
>site which has the info I need.
>>1. How easy is it to grow up large numbers of worms - say in the
>100,000 or more range?
>>2. Ia it possible to get this number of worms totally free from
>contaminating bacteria (for the experiment I have in mind they would
>need to have been feeding on bacteria for growth, which I assume is
>essential anyway) or does one have to go through a cycle of egg
>production?
>>Thanks for any help - I'm not a C. elegans researcher (yet!) hence the
>possible naivety of these questions.
>>Pete Lund
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