In-efficient Ligations? ANSWER!

Mark Youles my at mole.bio.cam.ac.uk
Mon Apr 17 03:56:30 EST 2000


Dear all,

Very many thanks for the responce to my initial post.  The common factor
which was in the majority of the answers, both here on the group and
directly sent to me via email (i.e. the UV light source), has now also
been tested.

I can further conclude from these tests that this WAS the causative agent
which had been preventing cloning.  When I directly compared the
transilluminator we are using with a different transilluminator (which is
used by many people and therefore I considered it 'safe') to both
visualise and subsequently cut-out the DNA bands requested, upon ligation
and transformation our transillumintor gave no colonies whereas the
alternative one gave a plate-full of re-ligation products (which were
SIGNIFICANTLY above any back-ground non-digested products).

Many thanks again for all your help.  Now I might actually be able to get
on with some real science and actually make my constructs!


Mark




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