3 replies to this topic

[ElHo]

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

[ursae]

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..

[mdfenko]

ursae, on Jul 18 2010, 09:50 PM, said:

y cant u heat becos of the urea?

urea will decompose and carbamylate the protein.

[redhill_2000]

try sonication. 30s.

good luck.