after adding phenol/chloroform to cells from culture and centrifugation, we get 3 layers. Upper 2 have RNA and DNA, while lower one (phenol) has proteins. can I extract these proteins? Which protocol Should I follow? I'm using the trizol reagent from qiagen.
Hans Matsson
After the Trizol extraction and separation of the protein fraction you can use spin-column filteres with e.g. 10kDa cut-off to get rid of any remaining DNA/RNA contaminants. Millipore and others sell these filters for E-tubes.
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Angelaalincy Maria Joseph
Yes you can. But you will have to filter and wash that layer to get rid of other impurities and contaminants. Then you can dialyse and obtain a partially purified protein.
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