Molecular cloning

Molecular cloning is a very important tool in the quest to understand gene function and regulation. Through molecular cloning it is possible to study individual genes in a controlled environment. Using molecular cloning it is possible to build complete libraries of fragments of DNA inserted into appropriate cloning vectors.

The in silico cloning process in CLC Main Workbench begins with the selection of sequences to be used:

        Toolbox | Cloning and Restriction Sites (Image cloningandrestrictionsites)| Cloning (Image cloning_16_n_p)

This will open a dialog where you can select the sequences containing the fragments you want to clone as well as sequences to be used as vector (figure 20.1).

Image cloningwizardstep1
Figure 20.1: Selecting one or more sequences containing the fragments you want to clone.

The CLC Main Workbench will now create a sequence list of the selected fragments and vector sequences (if you have selected both fragments and vectors) and open it in the cloning editor as shown in figure 20.2.

Image cloningviewinitial
Figure 20.2: Cloning editor.

When you save the cloning experiment, it is saved as a Sequence list. See Sequence lists for more information about sequence lists. If you need to open the list later for cloning work, simply switch to the Cloning (Image cloning_16_n_p) editor at the bottom of the view.

If you later in the process need additional sequences, you can easily add more sequences to the view. Just:

        right-click anywhere on the empty white area | Add Sequences