RNA ligation

Chris McDermott-Roe via methods%40net.bio.net (by cjmcdermottroe from googlemail.com)
Thu Nov 19 05:06:14 EST 2009


Hi,

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??

hope this makes sense

Chris

2009/11/19 Nick Theodorakis <nick.theodorakis from gmail.com>

> On Nov 18, 11:35 am, Chris McDermott-Roe
> <cjmcdermott... from googlemail.com> wrote:
> > Hi everyone,
> >
> > I hoping to ligate a synthesised oligonucleotide to the 5' end (uncapped
> but
> > not dephosphorylated) of an mRNA population.  Can anybody tell me the
> best
> > way of doing this?
> >
> > many thanks
> >
> > Chris
>
> Are they both single stranded? I think T4 RNA ligase will do that
> reaction.
>
> See:
> <http://www.neb.com/nebecomm/products/productM0204.asp>
>
> Nick
>
> --
> Nick Theodorakis
> nick_theodorakis from hotmail.com
> contact form:
> http://theodorakis.net/contact.html
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