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

Michael Witty mw132 at mole.bio.cam.ac.uk
Mon Nov 11 17:05:57 EST 2002


On 11 Nov 2002, 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.

I have always thought "yes", but it is inefficient.  Is it exonuclease V
which causes the low efficiency of linear DNA transformation in E coli?  I
also would appreciate expert opinion.  Mike.




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