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Vector & Insert???

Jack Leunissen jackl at caos.kun.nl
Sat Apr 6 07:34:24 EST 1996

Chris Barry wrote:
> Chris Barry wrote:
> > 
> > I have been playing around with the plasmid map on GCG. Could someone
> > tell me how I would go about:
> > 
> > 1) Digesting the insert and vector (will have stickey ends)
> > 2) Ligating the insert into the vector
> > 3) Displaying the new circular construct using the GCG plasmidmap
> > 
> > Thanks in advance for your time and patience.
> > 
> > Chris
> Ok! I am getting desparate now. If anyone has a clue to how to digest a
> sequence with one specific endonuclease, please let me know. I am using
> the graphical user interface to gcg wpi.
> Chris

OK Chris,
Hold your horses... First of all, it MIGHT HELP if you consulted the MANUAL,
like the rest of us do, before starting to shout!

Here's one way to do it:

ad 1) Digest the vector with MAP and the enzyme of your choice; from the
	output note the position of the cut-site.
ad 2) Use ASSEMBLE to construct the new sequence: vector (1 -- cutsite) +
	insert + vector (cutsite+1 -- end).
ad 3) Use program MAPSORT with the option "-PLASMID" or "-FRAGMENTS" to
	mark the position of the insert (so cut it with the previous enzyme
	again). Depending on whether you want to show the cutsites as
	tickmarks or as blocks, you choose either "-PLASMID" or "-FRAG".
	Use the resulting file as input for the PLASMIDMAP program.

Hope this helps you through Eastern.


       Jack A.M. Leunissen, PhD | CAOS/CAMM Center
       Email: jackl at caos.kun.nl | University of Nijmegen
       Tel. : +31 24 365 22 48  | Toernooiveld 1
       Fax  : +31 24 365 29 77  | 6525 ED Nijmegen, The Netherlands
                                | URL=http://www.caos.kun.nl/
     +------- CAOS/CAMM is the Dutch National Node in EMBnet -------+

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