Liquid Culture

Kevin O'Connell kfoconne at facstaff.wisc.edu
Tue Jul 11 16:44:49 EST 1995


In article <maske-1007951511370001 at mel.uoregon.edu>,
maske at molbio.uoregon.edu (MILO) wrote:

>    Has anyone had any luck growing C. elegans in liquid culture?  If so,
> or if anyone has grown up large quantities of worms by another method,
> please let me know.  Thanks.

I have had good luck using the following protocol:

1.  Grow OP50 to saturation in 2x YT medium.  

2.  Pellet and resuspend in 1/2 vol of the following medium:

            1L S-basal
            10 ml 1M K Citrate
            10 ml trace metals
            3 ml 1M CaCl2
            3 ml 1M MgSO4
            1 ml 10 mg/ml cholesterol

3.  Add worms that have been washed off of a uncontaminated plate.  I
inoculate a 500 ml culture with worms from a 100 mm plate.


4. Grow with constant shaking at 20-25 C .  It should take 4-5 days for
the culture to reach saturation.

I often find it necessary to add an antifungal agent such as nystatin.

Good Luck.

Kevin



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