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IanMc 10-27-2006 09:00 AM

Extraction of protein using trizol
 
Hi,

We are trying to fractionate gut samples into RNA/DNA/protein using the
protocols in the Trizol handbook. The procedure works OK but we are having
difficulty resuspending the protein pellet at the end of the process 1% SDS
or 1M Tris pH10.4 has not done the trick. We are about to try the urea/CHAPS
protocol. Does anyone have any other ideas on how to bring the pellet back
into solution?

TIA

Ian Mc



Nick Theodorakis 10-27-2006 03:36 PM

Extraction of protein using trizol
 

IanMc wrote:


No kidding. That means the procedure worked perfectly ;-)

Seriously, those pellets are very difficult to to resuspend. I use a
probe-type sonicator and sonicate about three or four times between
incubating the samples at 50C for an hour or two between sonications.
Basically, assume it takes all day, and plan to have something else
going on at the same time to keep yourself busy.

Nick

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