When I precipitate phages by PEG 8000, I got a very viscous pellets in one of these phages. Could someone tell me why the pellet is viscous? and does it effect any downstream analysis and does this tell anything about the phage that I am isolating? This is a novel phage.
Thank you.
Dear Tha, Your comment interested me as we have been using various PEGs to increase the viscosity of bacterial growth media (to look at the impact of bacterial swimming on localisation to the air-liquid interface). The recovery of an unexpectedly viscous phage pellet might be due to the lysis of phage which then release DNA (testable by DNAse treatment) or perhaps the interaction of PEG with the phage particles themselves (testable by diction with PEG-free buffer and re-pelleting). Regards, Andrew.
Hi Tha,
I just noticed this thread, and also get viscous PEG pellets when I resuspend in SM buffer. I usually add an equal volume of chloroform, vortex for ~30 seconds, then centrifuge at 5,000 x g for 15 minutes at 4C, transfer the aqueous layer to a new tube and repeat. The product is purified phage that is no longer viscous or cloudy with any cellular debris that got precipitated by PEG.
I hope that helps!
Nadia
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