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Lakia 05-14-2010 06:28 AM

Concentrating RNA
 
Hi all,

I have a few samples of RNA that I want to use for RACE PCR. The concentration in the samples is too low (around 30-60ng/microlitre) to make RACE cDNA at the moment but I think I can get a high enough concentration if I pool my samples (I have about 30 microlitres from several RNA extractions) and concentrate them down to about 8 microlitres. I am just wondering what the best method for concentrating the RNA. We have a speed vac in the building, but I think it only runs at ambient temperature at the best so I am a bit worried about degrading the RNA. Ive also considered ethanol precipitation but I'm worried about losing some RNA through that method. Ive had another suggestion that I can leave the sample in a fume hood and just let the water evaporate but that sounds a bit risky also. What is the safest method to concentrate the RNA?

Wandrews963 11-01-2010 10:16 PM

Re: Concentrating RNA
 
I don't know if this will help but in our lab when we concentrate RNA we use a Speed-Vac at 60 oC and dry for about 15 mins per 10 ul of RNA. The RNA is then fine for cDNA preparation and qPCR so I doubt the stability is being affected!

Hope that helps!
William

BTeske 11-02-2010 03:31 PM

Re: Concentrating RNA
 
The original poster has probably graduated by now...

Wandrews963 11-02-2010 04:51 PM

Re: Concentrating RNA
 
Oops did not even notice that! Tho it may help someone else so alls well!

BobMac 12-09-2010 04:56 PM

Re: Concentrating RNA
 
Quote:

Originally Posted by Wandrews963 (Post 423500)
I don't know if this will help but in our lab when we concentrate RNA we use a Speed-Vac at 60 oC and dry for about 15 mins per 10 ul of RNA. The RNA is then fine for cDNA preparation and qPCR so I doubt the stability is being affected!

Hope that helps!
William

This never occured to me to try. Definately worth remembering.

Thanks for the Tip!:clap2:


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