extra long ligation

[Kevin Urbanic]

Hi everybody,

I am having a problem ligating two 8kb DNA fragments together.  Each 
fragment was digested with the same enzyme.  I CIP-treat the vector and 
have attempted both chemical transformation and electrotransformation.  I 
have tried using I:V ratios of 1:1, 3:1, 5:1 with no success.

I am transforming with E. coli DH5alpha cells

Does anybody have any good suggestions or tricks that will help with this 
problem.

Thanks for your help.

Kevin



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Kevin Urbanic
Department of Microbiology
University of Guelph
Guelph, ON
N1G 2W1
Phone: (519) 824-4120 

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