T4 DNA Ligase: Oligo to Sticky End

Keith Rand Keith.Rand at molsci.csiro.au
Wed Nov 26 19:56:22 EST 1997

In article <65e15m$leb$1 at nargun.cc.uq.edu.au>, Paul R. Rohde
<P.Rohde at mailbox.uq.edu.au> wrote:

> T4 DNA Ligase joins blunt ended or cohesive-ended double stranded
> molecules but does not join single stranded molecules.
> My question:  Will T4 DNA Ligase join to a four base sticky end,a
> complementary oligonucleotide?  Yes, the oligo is single stranded,
> but the actual ligation site (or nick) is actually double stranded.
> This should work shouldn't it?  The oligo has been kinased to the 5'
> end.
>                          5'-gatcnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn...
>      3'-NNNNNNNNNNNNNNNNNNNNCTAGnnnnnnnnnnnnnnnnnnnnnnnnnnnnnn...
>                                ^^
>                              "nick"
>         (Oligo Capitals, ds Sticky End DNA lower case)

Yes, this should work. There is a paper somewhere that describes how you
can use short oligos to join 5' overhangs to 3' overhangs. If nobody else
replies and comes up with a suitable reference, and if you are interested
in looking at the paper, let me know and I'll search for it.

Keith Rand,  Sydney Australia

More information about the Methods mailing list