I've extracting total RNA from rat hearts with the standard TRIzol protocol.
Today, I altered the protocol to include a second CHCl3 extraction of the aqueous phase to more aggressively remove phenol. The aqueous phase as used in isopropanol ppt is a clear, slightly yellow solution.
After ppt, however, I noticed that the pellet is now peach-colored (It was transparent to white when I just extract with CHCl3 once), and 75% EtOH wash does not remove this colour. The resuspended RNA solution is also peach-coloured.
Does that mean the RNA has gone so unpure that I probably need to use cleanup columns? If so, what brand of column do you recommend?
Thanks!
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