Making of Blunt ends
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 ?
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.
Klaus Salger phone : +49 (0)89 5902 -502
Zoologisches Institut FAX : -450
AG Wetterauer e-mail: salger at zi.biologie.uni-muenchen.de
More information about the Methods