Ligation of short DNA fragments.

Zhongtang ztyu at yahoo.com
Mon Dec 16 14:02:02 EST 2002


Hi every one,

I want to construc concatemers of 40-bp dsDNA fragments by T4 ligation.
Each of my short DNA fragments has a 2-base overhang on either end. I
tried twice, but I did not get enough ligation. Judged from gel
electrophoresis, my ligation products ranged from dimers (a lot) to
about 400 bp (little). I need concatemers of >500 bp. The Technical
Support with Invitrogen told me that that will not happen. Does anyone
out there have similar experience, or suggestions? I got very large
concatemers in SAGE (with 4-base overhangs). Does the 2-base difference
make such a difference in ligation efficiency?

You suggestion and information will be very much appreciated.




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