Troubleshooting the "Unable to load recipe" error on the NovaSeq 6000

When attempting to set up a run on the NovaSeq 6000, some users have reported an error stating: "A problem occurred during run setup, unable to load recipes. to continue with run setup, reinstall the software suite". This error has also been reported to occasionally coincide with a second error stating: "A problem occurred during run setup. Reenter the run parameters or contact technical support".

This issue is most commonly caused by improper run formatting during run setup. Specifically, these errors will occur if a run has an index length of 0 bp for the i7/Index 1 while having an index length of ≥1 bp for the i5/Index 2. While Illumina sequencers do support single-index run formats, they do not support solely using the i5/Index 2 and setting the i7/Index 1 length to 0 bp.

Troubleshooting recommendations:

1. The best option is to construct the library or libraries to be single-indexed with only the i7/Index 1 instead of the i5/Index 2. A single-indexed run using only the i7/Index 1 is supported on all Illumina instruments.

2. If the error persists after making the run format change, power cycle the instrument and re-attempt the run setup.

   1. For further information, search for Knowledge Base article How to Power Cycle the NovaSeq 6000.

If the above troubleshooting steps do not resolve the issue, contact Illumina Technical Support with the following information:

• Intended run format (eg, 150 | 8 | 8 | 150, 100 | 8 | 0 | 100, etc.)

• NovaSeq Control Software Logs (C:\ProgramFile\Illumina\NovaSeq\NovaSeq Control Software\Logs)

• NovaSeq Run Setup Logs (C:\ProgramData*\Illumina\NovaSeq\NovaSeqTemp\RunSetupLogs[RunFolder])

*The ProgramData folder may be hidden; to un-hide files, select the View tab in the file explorer window and make sure “Hidden items” is checked.

• Screenshot of the recipes listed at C:\ProgramFile\Illumina\NovaSeq\NovaSeq Control Software\Recipes\NovaSeq6000

For any feedback or questions regarding this article (Illumina Knowledge Article #7943), contact Illumina Technical Support [e-mail].