RNA stability

Jan-Henner Wurmbach jhwurmbach at gmx.de
Fri Nov 19 15:00:05 EST 1999


On Fri, 19 Nov 1999 07:29:47 -0800, susanne
<srohrerNOsrSPAM at immv.unizh.ch.invalid> wrote:

>Hi
>
>I have been isolating total RNA from bacteria and storing it in dH2O at
>-80. Now I read on www.nwfsc.noaa.gov/protocols/ that it is a good idea
>to dissolve the RNA in formamide. It is said to keep it fully
>denatured, can be stored at 4°C overnight or indefinitely at -20. A
>first try Northern Blot looks different, possibly better than the
>earlier versions, I would even assume the RNA is "more denatured" than
>before, see below.
>Anyone tried this before? Does Formamide have to be of molecular
>biology grade, eg. deionized? what adverse effects would "bad"
>Formamide have?

Degraded Formamide is Formic Acid (German: Ameisensäure).
Probably reduces RNA Solubility and in the long run degrades RNA.

>Problem 2:
>Does anyone have experience with signals around the rRNA bands? could
>it be from interactions between mRNAs and rRNA, if it is not fully
>denatured? or could degradation products interact with rRNA?
>I have been detecting a major band just below the 16S rRNA band, and a
>minor fraction at the expected size (5kb). With the above formamide
>method, a strong 5kb band was detected, and the lower band was gone.
>
>Thanks for suggestions
>
>Susanne Rohrer, Uni Zurich
>
>
>
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