ligation problems. Help

M K Bennett pamkb at mail.bris.ac.uk
Tue Apr 25 06:48:24 EST 1995


Deborah Marshall (Marshall_D at a1.mscf.upenn.edu) wrote:
:    I am very frustrated because I haven't been able to get a 
: seemingly simple blunt-end, two-way ligation to work. My vector is
: 6.5 kb, with two homologous repeats (the LTRs in pLXSN), and the 
: insert is 3.5 kb (b-gal). I have tried transforming ligations into
: HB101 and XL1Blue, but keep obtaining colonies containing either 
: vector alone, or inexplicable deletions.


This may be a bit too obvious and Im NOT being sarcastic.  Remember that
after inserting b-gal the colonies to pick are the blue ones not the
white ones.  Even though there is no bacterial promoter b-gal inserted
like this will produce blue colonies.  

Mike 

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Mike Bennett D.Phil		|Scientists have odious manners, except when
Dept Pathology and Microbiology,|you prop up their theory; then you can borrow
University of Bristol, UK	|money from them--Mark Twain 1917 
m.k.bennett at bristol.ac.uk	|
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