Can a linearized Plasmid DNA ligate inside the E.coli?
hubahopp at gmx.de
Mon Nov 11 18:32:17 EST 2002
this result may be explained by uncut plasmid. However, you couldmix vector
and insert but not add ligase and transform tha t mix to see what happens.
In my experience, once when I tried it out of curiosity, it worked. My
explanation was intrinsic ligase activity.
At 07:20 PM 2002/11/11 -0000, Nagarajan Lakshmanan wrote:
>I have digested my plasmid with Xho1 and verified with agarose gel
>and gel purified a single band using Qiagen gel purification kit.
>When I transformed this DNA into DH10B, by electrophoration, I got
>several colonies. FYI, I do not dephosphorilate my vector. I think
>the linearized vector is ligating inside the E.coli cells. Could
>this be possible? I would appreciate your thought and experience on
>Lawrence Livermore National Lab.
>Biology & Biotechnology Research Program
>L-441, Building 366
>Livermore, CA 94550
>Phone: (925) 422 5783
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