Help wanted with blunt ligations (molecular biology)

Aart A. van Apeldoorn A.A.vanApeldoorn at
Thu Sep 21 11:38:37 EST 1995

This is a message for all you molecular biologists over there. I'm having
trouble with blunt ligations, i'm cloning a 1 Kb fragment into a 8.4 and a
4.5 Kb vector. Because of non comparable sticky ends i've blunted these
ends using klenow DNA polymerase. After blunting i ligate the fragment and
vector using T4-ligase in a volume of 10ul using 1 unit enzyme. After
transformation there are just a few or no colonies growing, my efficiency
is ok and it appears that there is no selfligation (after testing). Can
onyone give me some suggestions what is wrong here.    Thanks!

Aart A. van Apeldoorn
Dept. of Medical Physiology & Sports Medicine
Faculty of Medicine, Utrecht University
MFU-2 "Stratenum"
Universiteitsweg 100
3584 CG Utrecht
The Netherlands
E-mail: A.A.vanApeldoorn at

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