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I have isolated RNA from cell culture (T25 flask, mouse cells) with trizol. I've measured the sample on Nanodrop, and this is what I've got: A260 (10mm): 16.408 A260/A280: 1.98 656.3 ng/ul How is this possible? What could go wrong here? Thank you!

D.Novak
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  • What is your problem? The A260/280 ratio? The yield? I don't understand (actually see) the question. – Chris Apr 06 '17 at 12:38
  • The absorbance A260: 16.408 – D.Novak Apr 06 '17 at 12:47
  • The absorbance is directly proportional to the amount of nucleic acids (and proteins). Can you show the absorbance curve of your measurement? – Chris Apr 06 '17 at 12:56
  • no :( this is all i've got – D.Novak Apr 06 '17 at 13:09
  • Without any further data, we cannot give you any advice. But the A260 is not wrong, it shows that you have some RNA here. It is most likely still contaminated with Phenol though, this is what the high A260/280 ratio says. – Chris Apr 06 '17 at 14:07
  • Do you think RNeasy mini kit would help? Thank you very much for your help :) – D.Novak Apr 06 '17 at 14:27
  • I can't give you any advice as long as you are not sharing your protocol etc. Trizol in my experience gives fine RNA. – Chris Apr 06 '17 at 14:34
  • The 260/280 is almost 2, isn't that technically pure RNA? If the A260 readout alone is bothering you, read this article under Quantification of DNA – CKM Apr 06 '17 at 15:58
  • i used this protocol (RNA isolation procedure for cells) http://www.abcam.com/protocols/rna-isolation-protocol-cells-in-culture except I added 200uL chloroform, 500uL isopropanol and on the end I added 15uL DEPC treated water (because the last time there was small amount of cells). – D.Novak Apr 10 '17 at 12:16

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