I am doing a phenol/CH3 extraction with Life Technologies Totally RNA kit. I found that I had reached the bottom of the bottle of phenol that comes with the kit, and used another bottle that came from Fisher. I used the Fisher phenol for cleaning up a maxi-prep last week, so I didn't think there would be a problem.
However, when I added the Fisher phenol to the cell lysate, they did not form an interface or emulsion. I got thick, easily visible schlieren patterns in the tube which disappeared after heavy vortexing leaving a clear solution. Something about the Totally RNA denaturation solution makes it miscible with this phenol. I've been doing this for 7 years now and I haven't seen anything like this before. I'd like to recover these samples if I can. What's more likely to be successful? Addition of water or addition of phenol to break the interface? I have plenty of acid phenol from the kit, so I can do either. Or maybe addition of 50/50 h2o/phenol? _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
