5' biotinylation of RNA

Marc Lamphier lamphier at po.infosphere.or.jp
Tue Feb 20 10:19:57 EST 1996


In article <4g510m$d7u at news.xs4all.nl>, tvink at xs4all.nl (Tom Vink) wrote:

> Does somebody has a good protocol to label in vitro transcribed RNA at
> the 5' end with biotin. The only thing I could think of is ligating a
> biotinylated oligo to the end with T4 RNA ligase. Would it have to be
> a RNA oligo ?
> Tom Vink
> Department of Haematology
> University Hospital Utrecht
> The Netherlands

If you aren't worried about exactly where and how many biotin moleculaes
end up in the 5' region you could simply extend in the presence of a
limiting amount of biotinylated UTP, incorporating a few residues, and then
after a few munites finish the reaction by adding normal UTP. If you know
the sequence of the RNA you are synthesizing you can simply leave out one
of the nucleotides so that the reaction stalls until you add the normal
NTP.

-Marc 
University of Tokyo



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