Solubilizing RNA in water

Simon Mauch Simon.Mauch at uni-konstanz.de
Sun Jun 21 10:49:41 EST 1998


Hi Keith,
I added the "formamide"-RNA directly to the RT reaction. In my case I
used never more than 3 µl of formamide in a 30 µl RT reaction. (I use
Superscript II from Gibco.) The performance of the "formamide-RT" was
identical to a rection where the RNA was solubilized in water. 
So I had no problem with RT and formamide. 
The PCR also was no problem. 
Since formamide might influence the melting temperature of the primer
there might be a problem in some cases. 
In my reaction I am using 3 µl of the RT reaction in a 100 µl PCR so
its just about 0.3 µl of formamide..... which does not interfere with
primer binding...

Anyway, you could get rid of the formamide by normal ethanol
precipitation... But then you come back to the problem of RNA
solubilisation.....

I you want to meseaure the concentration by absorbance, don't forget
to ad formamide to the reference.

Hope this helps

Simon



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