RNA extraction from liver - (Aug/04/2005 )

Hi

I used Trizol-LS to extract RNA from liver. After homogenization, I added Chloroform and the left the mixture @RT for 15 minutes, then spin.

After spinning, there is no aqueous phase. Instead of aqueous phase, the top layer is white interphase.

Do you know what happen and what should I do?

Is the top layer aqueous? Put a drop of water in and see what it does. If the top layer is aqueous you can take it off, then I'd repeat the Trizol step. It just doesn't sound very clean.

Also make sure that you take the tubes out of the centrifuge immediately to avoid the interphase becoming dispersed. And make sure that you're spinning it enough. Don't laugh! I've put my samples in a centrifuge, gone for a cup of tea , came back and looked at my non-seperated samples. Guess who forgot to turn the centrifuge on???

Nicole

Is LS the trizol made for isolating from blood?? Dont know the difference between this and regular trizol, but may make a difference...

Liver has a TON!!! of lipids. Add more chloroform and re-centrifuge white interface may be lipid that was not well dissolved b/c not enough chloroform...

HTH
bec

thank you