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.



More information about the Methods mailing list