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pROTEIN & DNA

pROTEIN & DNA - Protocols and Methods Forum

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Old 07-26-2007, 08:47 AM
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Question pROTEIN & DNA



hello
is anyone know
how can we separate DNA from protein while extracting proteins from WBC by cell lysis method. Is it necessary to get rid of DNA?
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Old 07-27-2007, 02:32 AM
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Default Re: pROTEIN & DNA

I won't even say DNAse, because I don't think it's optimal for a protein mixture. But for most protein studies, gels, westerns, elisas, it isn't necessary nor desirable. Because you don't want your protein degrading while DNAse works.
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Old 07-27-2007, 10:36 AM
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Default Re: pROTEIN & DNA

if you just want your prots for crude prot work, ie, gel, immunoblotting and such then just lyse cells in lysis buffer (50ul 1M Tris pH 7.5, 4ul 0.5M EDTA, 40ul 10% Tx100 (Triton 100), 2ul 1M DTT,60ul 25xPI (protease inhibitors), 744ul SQ H2O. This is the one i use for adherent cells) by vortexing them in desired volume for 1 minute or so, then spin down 10min @ 14000rpm 4C and collect supernatant in fresh tube. That should be your protein, DNA is denatured and the lil that remains wont mess up your exp...

hope it helps
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