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Samy Terns
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Joined: May 27 2009
User offline. Last seen 2 years 28 weeks ago.

Does anyone know how I can prepare cell lysate from mix-staged worms using the glass douncer or any other method.  Thanks for your help!

CP expert
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Joined: Apr 3 2009
User offline. Last seen 1 year 1 week ago.
worm cell lysate

Pool worms and wash them in PBS by centrifugation, then rapidly freeze them in liquid N2. Grind them over a dry ice-ethanol bath using a mortar and pestle and add cold homogenization buffer (20 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM DTT, 1% Triton X-100, protease inhibitors just before use). You can then clear the lysate by centrifuging at 15,000 × g for 20 minutes at 4°C.
Alternatively, you can pool and wash worms as above, then resuspend worms in cold homogenization buffer and lyse by sonication on ice.  Finally, instead of sonicating you can pass the resuspended worms through a French press (1200 Kg/cm2) and then clear the lysate as above.

Anonymous
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