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 Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=3267