Try It Yourself (6 steps)

Digest and ligate can also be done by copying and pasting sequences. Please see Cloning by Copy and Paste.

Click here to pull in the data for this tutorial. The sequences pDusk and pDsRed2 will be added to a project called Training and opened for you.

We will do a simple digest and ligate using the Assembly Wizard by cloning the DsRed2 gene into pDusk.

1. Go to the Assembly Wizard at the bottom and click Create New Assembly. Select Digest and Ligate.Note: To access the Assembly Wizard, first open an inventory file.

2. Choose the bases for your backbone by going to pDusk. Click on the NotI site, hold shift, and click on the BamHI site. Right click and select Invert Selection. Your selection should look like the image below (6188 base pairs). In the Assembly Wizard, select Backbone and then Set from Selection.

3. Open the pDsRed2 plasmid by selecting it from the Workspace at the bottom left. Click on the Insert panel in the Assembly Wizard. Click on the BamHI site and then shift-click the NotI site (703 base pairs). Click Set from Selection. Your assembly should look like the following image.

4. Benchling noticed that the sticky ends of the two sequences did not match and showed error messages in the Assembly Wizard. To properly align the two sequences, click Reverse Orientation.

5. In the Wizard panel, name your assembly “pDusk-DsRed2 (from wizard)” and click Assemble.

6. When the new sequence opens, open the history panel on the right and click the Lineage tab. You’ll see lineage information about your assembly.

Did this answer your question?