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

SureGuide advanced wizard:

Add/Review Content

Upload gRNAs

The SureGuide advanced wizard allows you to upload your own gRNA sequences. You may want to use this method for adding content if you have a set of gRNAs that were designed with your own algorithms or were designed for a species that SureDesign does not support.

This method is also useful if you want to make edits to the gRNAs in an existing custom design. You can download the gRNAs from the existing design, make your edits, and then upload the edited gRNAs.

When you select to add content using the Upload gRNAs method, you provide a file containing the gRNA information, including sequences, and submit the file to SureDesign as a gRNA upload job. SureDesign uploads the gRNAs and saves them as a guidegroup. You receive an e-mail from Agilent SureDesign when the upload process is complete.

 NOTE  Prepare your gRNA upload file before you begin this step of the wizard. The maximum number of gRNAs permitted per file is 57,702 gRNAs for an amplified design and 244,038 gRNAs for an unamplified design. The file must be a tab-delimited text file (*.txt format) in a 4-column format. For upload purposes, create a compressed (*.zip format) version of the text file. Click here for further information on the file format requirements.

Complete the fields and selections in this step to designate the file you want to upload and set some of the parameters for the gRNAs.

Guidegroup Name

Type a name for the guidegroup that will be created from the uploaded gRNAs, or leave the default name in place. In a SureGuide design, the term guidegroup refers to a set of gRNA sequences. Designs are made up of one or more guidegroups.

File Name

Click Browse to select the gRNA upload file (in *.zip format). Alternatively, if you know the file path for the file, you can type it directly into the File Name field.

To submit the selected file for upload:

1. When you are ready to upload the file, click Next.
   
   A message box opens indicating the e-mail address where Agilent will contact you when SureDesign has finished uploading your gRNAs. If desired, you can enter additional e-mail addresses into the provided field in the message box.

2. Click OK in the message box to submit the file.
   
   SureDesign begins the process of uploading your gRNAs. The program first scans the file for formatting errors. If it detects a formatting error, SureDesign notifies you by e-mail. If it does not detect any formatting errors, SureDesign uploads the gRNAs into your SureDesign account.
   
   The wizard takes you to the Add/Review Content screen. The new guidegroup appears in the Guidegroup Summary table. The # Targets column of the table lists Processing until the probe upload job is complete. Click the refresh icon to see updates to this column.
   
   You receive an e-mail from Agilent SureDesign notifying you when your job is complete. In order to finalize the design, you must wait until your gRNA upload job is complete.