extra long ligation

Kevin Urbanic kurbanic at uoguelph.ca
Thu Apr 26 08:25:07 EST 2001

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 

Thanks for your help.



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


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