DNA reptation

Ferdinando Pucci brotizolam at libero.it
Sun Apr 4 15:54:37 EST 2004


Hi,
last week i was wondering why my 5' sticky 10kb DNA migrates bad on 0.5% agarose
gels. For bad i mean that to recover the whole band i should cut grams of gel.
Here is a description of the band under UV light: from a top point of view very
thin segments are visible, that make the tail of the band and get longer as the
migration take place. They're not visible on a photo of the gel. From the side a
parabolic shape is running, losing DNA from the top to the arms.

I read about DNA reptation of long fragments of DNA, 50kb and up. Given that
this linear DNA ends with GCGC (PauI sticky ends), that it tends strongly to
precipitate even in TE on ice, and that the only step after a good plasmid
isolation kit (qiagen, no affiliation) is a digestion, i'm asking if there are
long concatenamers that could affect the next ligation.
Thanks

-- 
Don't take life seriously, you'll never get out alive.



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