Making of Blunt ends

Klaus Salger salger at wap1.zi.biologie.uni-muenchen.de
Wed May 14 10:25:56 EST 1997


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,

I've always used T4-DNA-Pol with no problems. It's 3'-5'-exonuclease activity
is much higher than what you get with Klenow. Don't forget to put in dNTPs to
prevent the enzyme to remove more than just the 3'-protruding ends.

Cheers
  Klaus

--
Klaus Salger		phone : +49 (0)89 5902 	-502
Zoologisches Institut	FAX   :			-450
AG Wetterauer		e-mail: salger at zi.biologie.uni-muenchen.de
Luisenstr. 14
80333 Muenchen
Germany

BioLinks: http://www.zi.biologie.uni-muenchen.de/~salger/salger.html



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