Concentrating RNA - (May/13/2010 )
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?
As long as you don't mind losing some of the very small RNA, Zymo's RNA Clean and Concentrator columns work pretty well.
epibio on May 15 2010, 02:23 AM said:
i guess u can use any valid cut off centrifuge filters from any of the vendors... that way even the temperature u can control!!!
We normally just precipitate the RNA out of the solution using a carrier and resuspend in whatever volume we want.