Can a linearized Plasmid DNA ligate inside the E.coli?

Wolfgang Schechinger hubahopp at gmx.de
Mon Nov 11 18:32:17 EST 2002


Dear Naga, 

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.

Wolfgang


At 07:20 PM 2002/11/11 -0000, Nagarajan Lakshmanan wrote:
>Dear All,
>
>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 
>this.  Thanks.
>
>-- 
>Naga Lakshmanan
>Lawrence Livermore National Lab.
>Biology & Biotechnology Research Program
>L-441, Building 366
>Livermore, CA 94550
>Phone: (925) 422 5783
>---
>
>

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