I'm doing RNA extraction from plant cell suspensions, using TRIzol protocol. In the last steps, after the isopropanol precipitation and centrifugation, I get a pink pellet.
The color only appears after the addition of isopropanol (before the ispropanol, the solution is clear).
The 260/280 ratio is good, ~2, but I have contamination 260/230, low, i.e.
Hi Karla, I have this when I am extracting RNA by Trizol from plant material rich of anthocyanins and polyphenols and some of these molecules co-precipitate after isopropanol precipitation. If you wish you can get rid of the pink/purple color by re-precipitation with ethanol. But by my own experience this RNA is Ok for downstream application such as cDNA synthesis. Good luck!
Hi Karla,
There are 4 possible reasons causing the low A260/A230 ratio according to the attached NanoDrop bulletin, including the residual phenol contamination (see attached file).
If you think that the phenol is a possible problem, please see this website for possible methods to improve it:
http://www.protocol-online.org/biology-forums-2/posts/7291.html
Dear Karla,
Try replacing the final isopropanol precipitation with absolute ethanol and you should vbe all right.
Regards,
GCK
hello guys!!
I can not make it!! It is still pink!! i´m starting to think its something wrong with the TRizol reagent, what do you think?
Hmm, did you do a gel to see how your RNA looks like on it? If it is with good integrity go ahead with cDNA synthesis. For me it will be Ok even with "girl" pink color !
Good luck!
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