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