Destroy a Restriction Site

How can I simulate destruction of a restriction site in a circular plasmid by digestion and blunting (polishing)?

Specify the Restriction Site

To select the restriction site, click the enzyme name.

Open the Destroy Restriction Site Dialog

To open the Destroy Restriction Site dialog, click Actions → Restriction Cloning → Destroy Restriction Site..., or simply press the Delete key.

Preview the Product

The Destroy Restriction Site dialog will show the product preview. If the sticky ends were incompatible, they will be blunted automatically as shown in the overview at the bottom.

Blunting (or polishing) simulates enzymatic backfilling or removal of single-stranded overhangs - see Blunting.

In the above example, the enzyme T4 DNA polymerase could be used to remove the 3' overhangs generated by KpnI.

To view the settings, click the Vector tab at the top of the window, then review the blunting, enzyme selection, and selected site.

Specify the Bacterial Transformation Strain

To specify the bacterial transformation strain, click the Product tab, then click the Bacterial Transformation Strain menu to change the chosen strain or to edit the strains list.

Name the Product

Enter a name for the product, then click Clone.

View the Product History Colors

To turn on the product history colors in Map and Sequence views, click the "Show colors" button in the side toolbar. The red and blue colors represent where the restriction site was destroyed.

In Sequence view, the history colors illustrate more precisely where the restriction site was destroyed.

To illustrate in History view where the restriction site was destroyed, click the "Destroy Restriction Site" operation name.