Help! RNA precipitation emergency!

Nick Theodorakis nick_theodorakis at hotmail.com
Mon Mar 8 19:36:46 EST 2004


On 8 Mar 2004 12:01:17 -0800, nprigoda at netscape.net (N.P.) wrote:

>Hello everyone, I'm hoping that someone can help me solve this problem
>before my PI finds out about my error.
>I am extracting RNA from plant tissues using Trizol and a protocol
>that works very well. I pelleted my RNA and washed it.
>The problem is, instead of resuspending in water, I resuspended in 3M
>RNase-free NaAc.
>I now have RNA from 30 to 50 mg of tissue dissolved in 50µL of 3M
>NaAc, when I would normally have dissolved it in 50µL of water.
>Can I recover my RNA? If I follow the normal RNA precipitation
>protocol, then I should add 1mL of EtOH, but will this make my RNA too
>dilute to recover?
>Help!
>Nadia

If you have it in a 1.5 ml Eppie tube, add water to about 400 (you
want to dilute the salt some or will will fal out of solution) or so
microliters and 2 to 2.5 vol. ethanol, then chill and spin as usual.
Just call it an extra clean-up step to remove residual Trizol, rather
than a mistake ;-)

Nick
 

-- 
Nick Theodorakis
nick_theodorakis at hotmail.com
nicholas_theodorakis [at] urmc [dot] rochester [dot] edu 



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