When you have specified the insert and vector, and set up additional options as desired, you are ready to begin the cloning simulation:

  1. Press the Try It or Clone button in the bottom right corner of the Project window. The view switches from the Clone tab to the Summary tab.

  1. Review the three Summary sections: Fragments, Steps and Fragment Junctions. If the summary procedure does not meet your requirements, click Change to switch back to the Clone tab, where you can make changes. Otherwise, click Make It to proceed with the cloning simulation.

If simulation is successful, a new folder is added to Project tab of the Project window. Depending on the method used, this folder may contain a copy of each insert and entry clone, as well as the expected product that will be created after recombination occurs (i.e., the finished expression clone). Any item can be opened in the SeqBuilder Pro Document window by double-clicking on its name.

If simulation is unsuccessful, you may receive the error message “The vector is not digested by restriction enzyme(s).” This message may be seen when performing a cloning simulation where restriction enzyme cut sites were used to create the insert. Click Change to return to the Clone tab, then try again using a different insert.

  1. The Setup tab asks whether you wish to verify the clone. You can perform cloning verification at any time. If you have already prepared the clone in the laboratory and sequenced it, proceed to Step 6 (optional): Verify the clone. Otherwise, continue to the next step.
  1. Save the cloning project using File > Save Project. Then proceed to Step 7: View and print cloning results.

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