concentrate RNA - (Sep/18/2006 )
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.
glycogen is soluble in ethanol so you have to wash at least twice your sample.
sodium acetate should be removed also by ethanol 70%
There are commercially available (silica column based) kits for clean up and concentration of small amounts of RNA
the company starts with a Q
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.