freezing worms

rplas at RON.NKI.NL rplas at RON.NKI.NL
Fri Sep 23 15:40:02 EST 1994


>I am new to using C.elegans and have had difficulty freezing
>stocks of worms.  The N2 worm survived the protocol outlined
>in Wood's The Nematode C. elegans, but I've had no luck with
>the unc58 mutants.  Does anyone have a suggestion?
>
>Worm apprentice
>Rick Belval
>
>Rick Belval               | PO Box 5100     | 203-284-6378        
>belval at synapse.bms.com    | 5 Research Pkwy.| Fax:    
>Asst. Research Scientist  | Wallingford, CT | 203-284-6417 
>Bristol-Myers Squibb Co.  | 06492           | 
>Natural Products Research |                 |
--------------------------------------------

Having frozen 5,000 worm cultures, I would advise the following protocol,
which is easy:

Make regular 10 cm NGM plate, grow until all bacteria are completely gone, and worms
burrow into the agar. Then add a few ml of M9 buffer onto the plates, and shake overnight.
The burrowed worms will be washed out of the agar, and starve.
Take off the M9 plus worms, into a tube, let them sit for a while so that they sediment.
Take off most of sup, and add freezing solution. Freeze in a well isolated box in -70.


Ronald Plasterk



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