5'-end labeling of 8mer oligo?

Barry Henderson barry at aaRS
Mon Nov 14 15:27:48 EST 1994


pzhang (pzhang at cc.UManitoba.CA) wrote:
: Hi, netters,
: I posted my broblem a couple of days ago. Having got no reply, I'd like to try again.
: I want to radioactivly label 5'-end of a 8mer oligo. I am not sure how to separate labled oligo from the reaction (removal of ATP, ADP etc.) I want to use this oligo as a strand of an adapter. So, my second question is: If I use the labeled reaction directly( without any purification) to go ahead, what are the ligation results?

: Any ideas and suggestions are highly appreciated.


Why not just EtOH precipitate it and use it directly.  Your precipitation
efficiency may be a little low with such a short oligo but you should
be able to get enough back to work with.  Alternatively, gel purify it.
I routinely purify RNA oligos between 7 and 35 nts long on high percentage
(16%) denaturing acrylamide gels then extract the band you want (visualized
by UV shadowing or if it is labeled by autoradiography).

Barry Henderson



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