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Big Size of Plasmid Transformation


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#1 Isaiah

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Posted 08 April 2003 - 07:49 AM

Help!
I have trouble in transforming a big size of plasmid (about 40kb) into the competent cells. Since the recombinant plasmid is generated by homologous recombinant reaction, I can not use electroporation which needs large amount of DNA. I used normal competent cells (eg. Top 10, DH5alpha), heat shock method and got no colony after plating on LB plates with Amp as selective antibiotic.
Any good suggestion?

#2 zhenghj

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Posted 09 May 2003 - 05:18 AM

where do u get the idea that electroporation needs large amount of DNA?
the efficiency of heat shock is much lower than electrooration, and for such a
big plasmid, elctroportion is a practical way. Don't care about the amount of
ur DNA.Good luck.




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