Large plasmid cloning

Ferdinando Pucci brotizolam at libero.it
Sun Apr 4 07:12:17 EST 2004


On 4 Apr 2004 07:50:21 -0000
b8703126 at sinamail.com wrote:

> dephosphorylated by CIP, and purified by gel-extraction. The insert was also

Could you be more precise on which steps you do to dephosphorylate? In my little
experience i found that this step may have low yelds.

> purified by gel-extraction. And the ligation ratio i have tried was 2:1 and

Did you check at some point the presence of DNA?

> wrong with my cloning procedures? Have anyone of u experienced this kind of
> situation? Please give me some suggestions, thanks a lot.

I'm experiencing this with a plasmid of 15kb... Let's keep in contact! :-)

-- 
... I don't like FRANK SINATRA or his CHILDREN.



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