Self religation of double-cut plasmid vectors
AL R. PLUMMER
aplummer at super7.uccs.edu
Wed Jan 4 03:56:29 EST 1995
mahmoud (dbell at julian.uwo.ca) wrote:
: I was wondering in your hands, if you purify your double digested plasmid
: vector (any plasmid vector), and run a self religation experiment, what sort
: of transformation rate do you see? (how many colonies?) Of course I need to
: know how this rate is compared to the transformation rate of your non-cut
: plasmid control?
: In theory it should be zero, but...?
: Thanks in advance
Of course there are a lot of variables here, but in my hands there are always
a few transformants in a restricted, non-ligase treated plasmid prep. I
have always assumed that these were due to incompletely digested samples.
With ligase I generally obtain anywhere from 95-100% more 'true' transfor-
mants than without ligase treatment. In theory, you are correct, but
as with everything else in science-there are exceptions.
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