ligating single stranded DNA

Jim Kami jakami at ucdavis.edu
Mon Apr 24 12:12:46 EST 2000


Hi,
    Not only is it possible but very practical for a number of novel
cloning procedures. The "Band-Aid" oligo just needs to be annealed to
the fragment and transformed. I've been told that the ligase step is not
really necessary as the host cell will do that for you. A nice
application of this method is to use small oligos to create restriction
sites. The enzyme will only cut at the ds DNA region and then use a "
Band-Aid" oligo to religate the plasmid. A nice method when there are no
unique restriction sites in your region of interest. The only reference
I can think of offhand is one by Arrow and Dale 1985. I think it was in
Methods in Enzymology, but it's been so long I can't be sure.

Good luck,

Jim Kami
Department of Agronomy
Hunt Hall Rm 272
University of California
1 Shields Avenue
Davis, CA 95616-8515 USA
email: jakami at ucdavis.edu
Tel: (530) 752-9982
Fax: (530) 752-4361

"Why does Common Sense always seem to be the least common sense ?!?"

ed_taboada at my-deja.com wrote:

> Hi folks,
>
> I was wondering if it is possible to ligate two ssDNAs by having a
> linker fragment (or oligo) that hybridizes to both:
>
> ie.
>
>                        ---------
>          3'---------------5'3'--------------5'
>
> ideas ?
>
> tia,
>
> Ed
>
> Sent via Deja.com http://www.deja.com/
> Before you buy.
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