Transformation

Shilpashree Vinod Kumar sv56 at georgetown.edu
Mon Oct 2 15:29:15 EST 2006


   hey all,
   
   i am trying to do ligations with kpnI and HindIII at the ends in the
   insert. I am not able to see any insert after the
   transformation. Until ligation all the digestions and purifications
   have worked and the sizes of the insert and vector all look good. But
   when i screen colonies after transformation there will be no insert. i
   have tried different vector to insert ratios, i have tried ligations
   at 16 for 1hr, 2hr and also Over night. i am not understanding what is
   happening after ligation and transformation and where is the exact
   problem.
   
   Any suggestions??????
   
   Thanks
   
   Shilpa


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