T4 DNA Ligase: Oligo to Sticky End

[Chulho Kang]

> > 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)


 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

Dear fellows;

Yes, that is my paper. You can find  the short protocol at our company's
web site. Send your fax number, I will send the full copy of the
article.

Good Luck.

Biotechniques 1993 Oct;15(4):659-662  One-step insertion of
oligonucleotide linkers or adapters to DNA using unphosphorylated
oligonucleotides. 


Chulho Kang Ph.D.

Kang at hifax.com
Http://hifax.com