TA cloning

Mr Vid Mohan-Ram k9mhc at rfhsm.ac.uk
Thu May 18 22:47:48 EST 1995



hi,...

done the pcr - nice products,
 cut out the bands i wanted (200bp)
 ligated into pCRII vector (TA cloning kit, Invitrogen)....
did the transformation etc etc.....
....digested minipreps with ecori to check insert:-  get correct sized insert 
of 200bp but also an additional fragment of around 700bp (in all the 
minipreps)

this has happened to other people in the lab and on separate occasions,
two different lots of enzyme used, overnight or at least 5 hours digestion

anyone else had problems like this?

Vid
Vid Mohan-Ram
Dept. Anatomy and Developmental Biology
Royal Free Hospital School of Medicine,
Univesity of London,
UK.



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