purifying vectore, was Re: strange transformation results and
Michael L. Sullivan
mlsulliv at facstaff.wisc.edu
Sat Apr 27 12:54:43 EST 2002
>also, if the part of the MCS is still present in the ligation mixture, it
>will be religated back. To me it seems that the short DNA fragments ligate
>much more efficiently then the long ones (just a kinetic effect, much faster
>diffusion?). This will lead to more colonies on the control plate.
>Sequencing my constructs I have seen one with the right insert plus three
>MCS fragments ligated to each other. Since then I too always purify my
>vector on gel.
I too like to take the extra time to purify vector. I had a friend
in graduate school that thought this was a big waste of time. She
however, never counted in those 4 weeks she wasted figuring out that
her clone wasn't behaving as expected because it had polylinker
fragments as well as the intended insert.
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