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I added ethanol instead of chloroform to the cell suspension in Trizol. Can I still obtain my aqueous phase?

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  • $\begingroup$ How much ethanol did you add? $\endgroup$ – Chris Dec 18 '19 at 7:28
  • $\begingroup$ I added 100 uL of etanol $\endgroup$ – polonio210 Dec 18 '19 at 13:44
  • $\begingroup$ To which amount of sample? $\endgroup$ – Chris Dec 18 '19 at 14:11
  • $\begingroup$ to 500 uL of cell suspension in Trizol $\endgroup$ – polonio210 Dec 18 '19 at 16:08
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    $\begingroup$ @polonio210 Can you edit that information in the comments into the body of your question? $\endgroup$ – Bryan Krause Dec 18 '19 at 21:44
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My answer is based on my experience in the lab and something like a best guess - it may or may not work. But given the fact that added 100µl of ethanol to 500µl of total volume and since water and ethanol mix completely, I would give it a shot.

I don't think that the ethanol would change the polarity of the solution so much that phase separation is inhibited. The only thing that I can image is the contamination of the aqueous phase with traces of phenol, so I would probably clean up more carefully and do an additional NaOAC/ethanol precipitation at the end of the procedure. If you expect a small pellet with little nucleic acids, add some glycogen to get a better outcome there. I would appreciate it, if you could comment on your experience (and the outcome) when you try to save you sample.

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    $\begingroup$ Thanks Chris, Oh yes, I will let you know! $\endgroup$ – polonio210 Dec 19 '19 at 16:25

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