T4 DNA Ligase: Oligo to Sticky End
kang at hifax.com
Mon Dec 8 06:01:13 EST 1997
> > 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
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
Biotechniques 1993 Oct;15(4):659-662 One-step insertion of
oligonucleotide linkers or adapters to DNA using unphosphorylated
Chulho Kang Ph.D.
Kang at hifax.com
More information about the Methods