editing problem with large constructs

Stuart Kuhstoss sk at lilly.com
Thu Dec 15 12:10:15 EST 1994


In Article <1994Dec13.092837.5764 at ucbeh>, converrl at ucbeh.san.uc.edu wrote:
>     I am in desparate need of suggestions for constructing large plasmid
>clones.  Presently, I am attempting to build a 12 kb construct in bluescript by
>cloning a 2 kb blunt5'/sticky3' fragment into a 10 kb bluescipt fragment wich
>has identical ends.  I am getting lots of transformants, but they are all
>severely edited. The bacterial strain I am using is XL-1 Blue.  I have also
>tried SURE cells, but I get the same results.  I would greatly appreciate any
>helpful suggestions. Please E-mail them to me!!
>
>                           Thanx!
>
>                           Richard 
+++++++++++++++++++++++++++++
You might try DH10B from BRL.  I've had good luck with these as a host for
numerous constructs, including some of around 20 kb and containing inserts
with around 70% GC.

Stu Kuhstoss



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