Ligation of short DNA fragments.
hubahopp at gmx.de
Mon Dec 16 18:30:21 EST 2002
You also should use a ligation buffer that contains some viscuous stuff
like PEG (e.g. the buffer from LifeTechnologies).
At 02:02 PM 2002/12/16 -0500, Zhongtang wrote:
>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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