Palindrome and ligations

Francois Nantel Nantel at
Tue Jul 25 06:17:04 EST 1995

For the past few month or so, I have been desperatly trying to do a
ligation in which a 3.2 kb blunt-sticky fragment is inserted into a 4.8
kb vector. Up to now, all I have are empty plates. I tried a lot of
different protocols and purifications to no avail.

Recently, I began to wonder if my problem had to do with the nature of
the insert. In fact, this insert contains the loxP sequence very close to
the edges. The loxP sequence is a 32 bp palindrome used for recombination
experiments. I am wondering if such a large structure can interfere with
the ligation reaction and, if so, how could I correct it?

Any hints and comments would be greatly appreciated.

Francois Nantel
nantel at

Strasbourg, France

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