Lysing worms for biochemistry

LJAC at VMS.CIS.PITT.EDU LJAC at VMS.CIS.PITT.EDU
Tue Sep 16 11:23:18 EST 1997


>I am interested in getting worm lysate for co-immunoprecipitation and
>the like, and I am wondering if anyone has suggestions on preparing the
>worms.  Is freeze-thawing adequate, or should we add sonication or other
>mechanical disruption?
>
>Garth Patterson
>Ruvkun Lab
>

Depends on what scale you have in mind. For very small samples, we find that 6
freeze-thaw cycles (perhaps with a small amount of NP-40) is adequate for
nearly all purposes. At intermediate scale, sonication is fine, but is
extremely difficult to do with high reproducibility if quantitation is desired.
At larger scale (>3 ml suspension), we like the French press, which makes very
reproducible high-quality extracts.

Lew Jacobson
Biological Sciences
Univ. of Pittsburgh
Pittsburgh PA 15260
412-624-4637
LJAC at vms.cis.pitt.edu





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