RNA precipitation - (Apr/28/2006 )
I want to concentrate a sample of RNA. I think that I have to precipitate it and then, resuspend it with a lower volum of water. How can I do it? Can you tell me a protocol?
add 1µl of 100mg/ml glycogen, mix gently to homogenize, add then 10volumes of cold butanol. Vortex well to homognize.
Spin 20 000g for 10'. Wash twice with 1ml ethoh 70% and resuspend in less volume.
Fred you don't need to add glycogen if you use butanol as it works by dehydration rather than condensation like ethanol. Also it doesn't need to be cold - RT works fine.
In our experiment we are using glycogen and ammonium acetate or sodium acetate for precepetation and incubate for 30min at 4C and spin its and dry pellete and resusped in low volume of DEPC treated water.
all the best.
i use glycogen as a carrier for low quantities of RNA, as it may not be all pelleted sometimes.
My experience is that you don't need to use butanol cold, however, I don't know if you need to add a carrier for low conc. of RNA. I guess it won't hurt and it will give you a pellet you can see.