[email protected] (DK) wrote: >is the best way of cleaning up such reactions to remove salts and >nucleotides? Without losing the precious RNA, of course.
I would go for ammonium acetate to remove the free nucleotides. tRNA are used as a carrier for small amounts of DNA, so I wouldn't worry too much. There is a interesting paper about it from the Bethesda Research Laboratories: Crouse J, Amorese D (1987). "Ethanol Precipitation: Ammonium Acetate as an Alternative to Sodium Acetate". Focus 9 (2): 3-5. The PDF can be found here: http://bio.wayne.edu/profhtml/Cunningham/private/privatedocuments/EtOH.pdf (and probably also somewhere on the Life Technologies website). >For EtOH pption: How long to keep on ice? (Is low temp even >needed; for DNA I find that it is not beneficial at all). What cation There is another interesting paper also from BRL, which finds surprisingly little effect of cooling in the precipitation at all: Zeugin JA, Hartley JL (1985). "Ethanol Precipitation of DNA". Focus 7 (4): 1-2. http://www.lifetechnologies.com/content/dam/LifeTech/migration/en/filelibrary/pdf/focus.par.56415.file.dat/focus%20volume%207%20issue%204.pdf Christian -- X-no-Sig: yes _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
