Unstable insert

Bas Jansen bas at nospam.nl
Tue Feb 8 07:13:59 EST 2000


Rafael Maldonado heeft geschreven in bericht <389FDEA0.605D9A2A at ua.es>...
>
>
>panapat at netscape.net wrote:


[Problems with cloning a 10kb insert in pUC18]

>Besides the recommenddations by Jered, you should use a different
>vector. pUC's are not designed to carry those big inserts you are
>cloning. A lower copy number vector is more suitable.


To my knowledge, pUC18 with inserts of 10kb should propagate just fine in
DH5alpha. If the insert is lethal or unstable, the original poster could try
pUC19. I have in the past solved this kind of cloning problems by simply
reversing the insert by using pUC19 instead of pUC18, or Bluescript KS
instead of SK.

Bas
basjhj at baserv.uci.kun.nl








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