I have an issue. I isolated RNA from cortex a few days ago. For isolation of RNA i used this procedure:

I homogenizised my sample (mouse cortex) with Trizol. Then to the sample add chloroform, shooke and centrifugate it. After this part i took away the aquatic phase with RNA from mixture and add it to the eppendorf tube with beforehead prepared isopropyl alcohol with glykogen. Again i shook and centrifugate it. In this part i removed isopropyl and let only RNA pelet. Afterwards i did same thing with 75% ethanole. Last but not least i dissolved the pellet with DEPC, checked its concentration on nanodrop.

From nanodrop i saw that one of the sample had been contaminate with guanidine isothiocyanete. So i just desided to repeat steps od my isolation from isopropyl alcohol. To increase precipitation of RNA and increase chance od decontamination i decided to let RNA in isopropyl alcohol precipitate in -80 °C freezer for a night. Unfortunately i fail to remember it untill now.

My question is: Will it affect any integrity of my RNA sample?

  • 2
    $\begingroup$ It would generally not be an issue to leave RNA in alcohol but you should clarify your question. What do you mean by "let it clean"? Why are you doing it after the extraction and quantification? What was the concentration of alcohol? $\endgroup$ – WYSIWYG Dec 4 '19 at 10:35
  • $\begingroup$ WYSIWYG i try at least write down the steps i did. Today i was at lab but i forgot check the concentration. But i usually use all chemicals in proportion to volume of trizol. So the isopropyl alkohol is from bought storage bottle. $\endgroup$ – L.Diago Dec 4 '19 at 21:24

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