Blunt ending??

Chris Boyd chrisb at
Tue Jun 9 06:19:28 EST 1998

jalali at 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  | on behalf of  /  Western General Hospital, \   Edinburgh, EH4 2XU, SCOTLAND

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