protocol for purification of oligonucleotide

Giovanni Maga maga at vetbio.unizh.ch
Mon Jan 23 18:31:33 EST 1995


In article <95018.163834LIUG at QUCDN.QueensU.CA>, <LIUG at QUCDN.QueensU.CA>
wrote:

> I am looking for a protocol for purification of oligonucleotides (15-40 bases)
> after the kinase reaction. If any one has a fast protocol, please send me a
> E-mail.
>             Ge Liu, Dept. of Microbiology, Queen' university
>             Liug at qucdn.queensu.ca

I guess NAP or NICK columns from Pharmacia are the best choice (they
provide customers also with detailed protocols). If you want to try
yourself (just for the sake of curiosity) I suggest you to fill up a
pasteur pipette with G-25 Sephadex, load your labeling reaction, elute with
TE. If you follow the radioactivity with a geiger, you will see a first
peak in the void volume (oligo) and a second peak after (ATP). Take your
oligo and precipitate it as you want. (A hint: probably you will find
useful to put a little bit of glass wool on the bottom of your pipette
before pouring in the G-25).
Good Luck.



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