TA cloning vector w/ ddTTP (ref!)

[Helge Weissig]

>>how come that you get ligation to a ddTTP tailed DNA strand?
>
>>puzzled,
>>helge

cdolphin at crc.ac.uk (Dr. C. Dolphin) wrote:
>>
>Dear Helge, obviously you don't get ligation on that strand but you will on
>the other one at that position, ie. you get a sort of double nicked
>(half-ligated)
>plasmid. This seems to be fine for transfection and then the bugs produce super
>-coiled. We've used Taq to do the same thing but with dTTP rather than ddTTP
>and this also works but how efficient is this activity of Taq ? 
>
>Colin
>
>Colin Dolphin                           Email: c.t.dolphin at qmw.ac.uk
>Biochemistry                            Tel: (44) 71 975 5555
>Queen Mary & Westfield College  FAX: (44) 81 983 0531
>London E1 4NS, UK


Dear Colin,
in the meantime I even found the reference for it :)

                i.e. Holton and Graham, NAR 19: 1156 (1991)

and looking at it now, I realized that it is just 2 pages after the one I
have been using so far.

cheers,
helge



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Helge Weissig      

La Jolla Cancer Research Foundation             (619) 455 6480 x253
10901 N. Torrey Pines Rd.                  FAX: (619) 455 0181
La Jolla, CA 92034                      e-mail: helgew at ljcrf.edu
USA

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