cdna cloning with stratagene cdna cloning kit
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.
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