Troubleshooting "GenerateFASTQ 500 Internal Server Error" in Local Run Manager hosted on the iSeq 100

Users may encounter this Internal Server Error after updating the iSeq 100 Software Suite to v3.1, which also updates Local Run Manager Framework to v4.2.0. Upon installation of the GenerateFASTQ v3.1.0.35 module, users may not be able to access GenerateFASTQ through Local Run Manager (LRM), which then prevents the user from creating a new run for sequencing.

This error may be caused by installing the incorrect module version or unexpected failures during software suite update.

Troubleshooting steps:

There are two options that may resolve this issue:

Option 1: Uninstall and Reinstall Module and Software Suite

Option 2: Restore system to factory settings (Uninstalls software and clears contents from C:\ drive)

NOTE: Whichever option is chosen, Illumina highly recommends re-downloading the iSeq System Software v3.1 and LRM module installers.

Option 1:

Uninstall and reinstall module and software in the following order, as per the software release notes:

1. Uninstall Generate FASTQ module

2. Uninstall iSeq 100 v3.1 Software Suite (reboot required at the end)

3. Install iSeq 100 v3.1 Software Suite (reboot required at the end)

4. Access Local Run Manager via Chromium Browser (https://localhost) and log into LRM Admin account to confirm it does not require re-configuration

5. Install Generate FASTQ module (it may take up to 30 minutes)

Option 2:

Re-install software in the following order, as per release notes (Refer to Restore to Factory Settings section of the iSeq 100 Sequencing System Guide):

1. Install iSeq 100 v3.1 Software Suite (reboot required at the end)

2. Access Local Run Manager via Chromium Browser (https://localhost) and configure new LRM Admin account by resetting password and completing security questions and answers

3. Install Generate FASTQ module (may take up to 30 minutes)

For any feedback or questions regarding this article (Illumina Knowledge Article #9035), contact Illumina Technical Support [e-mail].