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direct lysis of cell pellet


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#1 ElHo

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Posted 15 July 2010 - 01:22 AM

Hello all,

I tried to directly lyse my cell pellets in a buffer containing 8M urea by incubating at RT for 1h. But when I tried to load my samples on SDS-PAGE, the solution was far too sticky. In my opinion this is due to DNA.
I have already tried centrifugation to pellet the DNA and passing the solution through a needle and both did not work. I do not want to heat my samples because of the urea. Any additional ideas how to get rid of the DNA?

Thanks in advance

Ellis

#2 ursae

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Posted 18 July 2010 - 05:50 PM

firstly, y cant u heat becos of the urea?

have u tried using QIAgen shredder? It will help to get rid of the DNA but its kinda time-consuming though..

#3 mdfenko

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Posted 19 July 2010 - 09:40 AM

y cant u heat becos of the urea?

urea will decompose and carbamylate the protein.
talent does what it can
genius does what it must
i do what i get paid to do

#4 redhill_2000

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Posted 24 October 2010 - 02:34 AM

try sonication. 30s.

good luck.




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