Protocol Online logo
Top : Forum Archives: : Molecular Biology

concentrate RNA - (Sep/18/2006 )

Hi Everybody!!!

I have to concentrate RNA to perform a Retrotranscription and later on Realtime-time PCR. I know that with such few amount (1 ug of RNA) , I could precipitate with glycogen and Ammonium Acetate. I'm wondering if these reagents could interfere with RT or RT-PCR. Could you recommend me some kit that works well? I have to concentrate up to 10 uL.

Thank you.

-sus12-

glycogen is soluble in ethanol so you have to wash at least twice your sample.
sodium acetate should be removed also by ethanol 70%

-fred_33-

There are commercially available (silica column based) kits for clean up and concentration of small amounts of RNA

the company starts with a Q wink.gif

-John Buckels-

oh and i think of a cheap method...
Precipitate with butanol (10volumes is good)
Add BuOH, vortex, spin 5' wash 2x with etoh 70 dry and resuspend in smaller volume.

-fred_33-