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chloroform extraction---become dark yellow


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#1 jenny0827

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Posted 06 May 2010 - 12:06 PM

Today when I did phenol/chloroform extraction for the ChIP DNA, I found something strange. After adding chloroform to the aquar phase which extracted from phenol/chloroform/Isoamylalcohol, the organic phase turned to dark yellow...I found maybe due to chloroform is old, because the chloroform I used was a bit yellow...phenol/chlo/Isoamyl should be ok, it was a commercial one, the color is still light yellow...Is there anyone knows what is the dark yellow thing, will it break the DNA???

#2 hobglobin

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Posted 06 May 2010 - 12:42 PM

Today when I did phenol/chloroform extraction for the ChIP DNA, I found something strange. After adding chloroform to the aquar phase which extracted from phenol/chloroform/Isoamylalcohol, the organic phase turned to dark yellow...I found maybe due to chloroform is old, because the chloroform I used was a bit yellow...phenol/chlo/Isoamyl should be ok, it was a commercial one, the color is still light yellow...Is there anyone knows what is the dark yellow thing, will it break the DNA???

normally the phenol can turn yellow if old and oxidised...it's not recommended to use it then...

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#3 pDNA

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Posted 06 May 2010 - 02:07 PM

Oxidation of the phenol produces a pink/brown compound, and this compound will cause nicking of your DNA and degradation of your RNA.

Regards,
p




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