Making of Blunt ends

Martin Offterdinger a8803349 at unet.univie.ac.at
Wed May 14 06:21:34 EST 1997


On 14 May 1997 11:27:39 +0100, Claus Rodemer
<c.rodemer at dkfz-heidelberg.de> wrote:

>Hi all,
>which is the best way to make blunt ends ? I have cut  the DNA with a enzym
>that  makes 3' protruding ends and I want to make blunt ends out of it.
>
>Is Klenow Ok, or T4 DNA-Pol ?  Or is it possible with S1 Nuclease,too?
>
>Another thing:
>I tested a cloning kit, and with this kit there is a "polishing reaction"
>provided (cloned Pfu Pol) .The manual said that this makes blunt ends ,too.
>Well I am still testing ... anybody knows something about this ?
>
>Bye,
>Claus
>--
>
>
>------------------------------------------------------------------------
>Claus Rodemer  German Cancer Research Center  INF 280
>Div.Biophysics of Macromolecules  AG. Gene Regulation and DNA Topology
>D-69120 Heidelberg  E-Mail: C.Rodemer at dkfz-heidelberg.de
>WWW:http://www.rzuser.uni-heidelberg.de/~crodemer/
>------------------------------------------------------------------------
>
>
S1 nuclease should not be used because there is a tendency to loose
also base-paired nucleotides, when overdigestion is accidentally
performed, I always use Klenow with good success.
Martin



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