Assemble guide RNAs (gRNAs) into plasmids

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In this tutorial, we’ll assemble a few guide RNAs into plasmids. Before you start, follow along with the Design guide RNAs tutorial to create some guides for the Brca2 gene. We'll use the guides you created in that tutorial in this example.
If you don’t already have the CRISPR analysis open, open it by clicking the Genome Engineering panel on the right and choosing the saved CRISPR design labeled Exon 2/3.

Sort by on-target score and select the first 4 guides. Click Assemble.

Benchling provides a list of plasmids that are commonly used for gRNA assembly. Select the plasmid LENTICRISPRV2 (52961) as your vector source and click Next.

Click on Choose Folder to save your assembly in a specific folder. Once the window opens, click on Create new folder and label the new folder “BRAC2 CRISPR Plasmids”. Click Create and Select.

Click Assemble to assemble your gRNAs into the plasmid.

From the message that appears, create a notebook entry of your assembled gRNAs by clicking create a notebook entry.

View the entry, along with the linked protocol, that is added to your notebook in Training. You can edit this protocol to customize it for your particular experiment. For example, scroll down to Table 2 and change the number of pairs to “3”. Notice that Table 3 is linked to this cell and that the Master Mix (uL) column updated.


You can also print your entry by clicking on the “print” icon on the top and choosing Print Entry.

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