The functionality described in this topic is only available when you mark Show Advanced Options.

SureGuide advanced wizard:

Define Design

In this step, complete the fields described below to define the design.

Design Name

Type a name for your design into the field. Alphanumeric characters, hyphens, underscores, and spaces are permitted. The name must be unique within your workgroup.

Species

Specify the species of the targets. The default selection is H. sapiens.

To change the selection, click Select. The Select Species dialog box opens, allowing you to select from a list of available species.

Build

If multiple genome builds are available for the selected species, select the desired genome build in the provided drop-down list.

If no drop-down list for build selection is provided, that indicates that only one genome build is currently available for the selected species. That build is indicated below the Species field.

 NOTE  SureDesign allows you to upload custom genomes to use when creating SureGuide designs. Click Manage Genomes to view the available custom genomes and upload new genome files.

CRISPR Application

Select the appropriate CRISPR application for the design.

·        CRISPRa - CRISPR activation (for up-regulation of a target)

·        CRISPRi - CRISPR interference (for down-regulation of a target)

·        CRISPRko - CRISPR gene knockout (for knockout of a target)

Category

Specify if you want the gRNA library amplified or unamplified. Amplified libraries are composed of linear DNA fragments with end-sequences that make the oligos suitable for cloning into Agilent's pSGLenti vector backbone. Unamplified libraries are also composed of linear DNA fragments, but the end-sequences flanking the gRNA-encoding region are fully customizable. In order to create a plasmid library from an unamplified gRNA library, you must first PCR-amplify the gRNA library and then clone the amplicons into a suitable vector.  

Create In

Specify the folder in which you want to save this design. The default selection is the top-level folder for your workgroup.

To change the selection, click Select to open the Select Folder dialog box, and mark the folder in which you want to save the new design. This dialog box lists the available folders within your workgroup and, if you are a member of any collaborations, lists the collaboration folders to which you have access. (If you later decide you want to change the folder location of the design, you can move it to another folder.)

Click Next to advance to the Add/Review Content step.