X-Message-Number: 3267
Date: Fri, 14 Oct 1994 18:40:34 -0400 (EDT)
From: 
Subject: SCI.CRYONICS Worm Apprentice Freeze

Date sent:  14-OCT-1994 18:37:19  

>Sender: 
>>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               | PO Box 5100     | 203-284-6378
>+---------------------------------------------------------------------------
>>    | 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.
>
>
>REPOSTED by Jan (John) Coetzee

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