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TA cloning vector

louhelainen at cc.helsinki.fi louhelainen at cc.helsinki.fi
Sat Aug 7 13:58:03 EST 1993


In article <CBCrG9.9nI at ccu.umanitoba.ca>, browns at ccu.umanitoba.ca (Stuart Brown) writes:
 
> I have had mixed success with the Invitrogen kit.  The secret seems to 
> be choosing the right colonies off of your kanamycin plate.  Not the
> white ones, not the blue ones, but the sort of whitish ones with tiny
> blue dots in the center.  I don't know why this is, but there is some
> mumbling in the kit's instructions about out of frame religation of the
> vector forming white "background" colonies.  

  Sounds like you're getting light-blue colonies. Maybe your insert has an
  initiation codon and it lacks in-frame termination codon ? 
  Sometimes you get false positive LB-colonies, if there has been a base pair 
  deletion during the linearization/religation step. 


  Jari Louhelainen
  University of Helsinki
  Department of General Microbiology
  FINLAND



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