RNA ligation

Aawara Chowdhury via methods%40net.bio.net (by aawara from pontiff-playground.org)
Fri Nov 20 06:19:26 EST 2009


In <63e4a4c6-9734-4886-baba-dab9a41e76b7 from a21g2000yqc.googlegroups.com>,
 Nick Theodorakis <nick.theodorakis from gmail.com> wrote:

>> Yes, they'll both be single stranded. ?One will be the mRNA that I want to
>> tag (at the 5' end), the other will be a ss oligo. ?My concern is that the
>> oligo may ligate to the 3' end of the mRNA but I guess this will depend on
>> how I have the oligo modified? ?Is it sensible to assume that if it (the
>> oligo) is 3' hydroxylated but NOT 5' phosphorylated, then it will bond with
>> the exposed 5' phosphate on the mRNA and NOT the 3' OH??
>
> That sounds sensible. If the oligo has no terminal phosphates and the
> RNA is 5' phosphorylated, then the only reactions that should happen
> are ligation of the oligo to the 5'end of the RNA .. [deleted]..

Won't work for mRNA - there is no free 5'phosphate.

AC
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