Blunt ending??

Chris Boyd chrisb at hgu.mrc.ac.uk
Tue Jun 9 06:19:28 EST 1998


jalali at my-dejanews.com wrote:
: I need to blunt end a linearized plasmid with a 5'
: overhang of 2 bases, as well as a fragment of DNA with a 5' overhang.
: The strategy I'm considering at the moment is digestion with Mung-bean
: nuclease. Any thoughts?

Everyone else seems to recommend Klenow + dNTPs.  For reasons outlined
in (1) I would recommend T4 DNA polymerase + dNTPs for all routine
blunt-ending.  It works for me, at least...

1  Costa, G. L. and Weiner, M. P. (1994) `Polishing with T4 or Pfu
   polymerase increases the efficiency of cloning of PCR fragments.'
   Nucleic Acids Res., 22, 2423


Best wishes,
--
Chris Boyd                      | from, but not \ MRC Human Genetics Unit,
Christopher.Boyd at hgu.mrc.ac.uk  | on behalf of  /  Western General Hospital,
http://www.hgu.mrc.ac.uk/Users/Christopher.Boyd \   Edinburgh, EH4 2XU, SCOTLAND



More information about the Methods mailing list