cdna cloning with stratagene cdna cloning kit

Andrew Mendelsohn mendelsohn at frodo.mgh.harvard.edu
Thu Jul 25 13:26:00 EST 1991


In article <24JUL199116174558 at frodo.mgh.harvard.edu>, mendelsohn at frodo.mgh.harvard.edu (Andrew Mendelsohn) writes...
My original posting was trunscated here is the full posting  (i hope)
:  We have been having trouble with the stratagene cDNA directional 
cloning kit and with our own version of their reagents.  We have not 
obtained many inserts corresponding to cDNA with their  
>strategy.  We see stimulation of plaques or  colonies after adding the 
>cDNA to the vector for ligation, but do not get many cDNA inserts.  
>We probably are getting some of the adaptor/primer in our false positives.
>Has anyone had trouble with getting the methylated cdna to clone in 
>the SURE strain.  Any similar troubles removing adaptor-- we use both
>spin columns and salt gradients to fractionate our cDNA before cloning.
> 
Any help would be appreciated.
andy



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