Any idea about the protocol to separate polysomes without using density gradient fractionation system?

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Karthik Raman

I did a quick search on SciGine and found this method for polysome extraction using a sucrose cushion. Perhaps it will help.

http://scigine.com/method.php?ID=115975

http://scigine.com/method.php?ID=154502

Brian Bower

That can't actually be 10 cells/ml, but what is the real quantity for the protocol?

I'd like to test this method vs setting up the sucrose gradient technique.

Okay, there it is, in the original paper. It's 1e8 cells/ml. Holy macaroni.That would be just about a solid pellet of cells unless I'm mis-remembering my cell culture days. (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2045085/)

Brian, yes you are correct. The scigine website seems to indicate wrong cell counts because of the ^ symbol.