Troubleshooting fluidics automation error on the NextSeq 500/550

Possible causes of the fluidics automation error on the NextSeq 500/550 include:

• Communication issue between the hardware and software.

• The waste/spent reagent container is not fully inserted.

• A hardware component issue.

These can occur during NextSeq 500/550 control software initialization, during a run, or during a wash.

Example notification in NextSeq 500/550 control software:

Descriptions of Fluidics Automation Errors that can be observed in the control software:

• 1: Operation home failed due to flowcell_z not being able to move to flowcell_z_home=1

• 2: Operation home failed due to flowcell_z not being able to move to flowcell_z_home=0

• 3: Failed to move flowcell_z to down

• 6: Move flow cell to out failed due to flow cell home not being 0

• 7: Failed to move flowcell_x (FlowCell Loading Mechanism)

• 12: Failed to move flowcell_align to out

• 20: Failed to move sipper_y to pick

• 22: Move reagent z to down failed due to reagent tray up not being 0

• 24: Operation home failed due to reagent_z not being able to move to reagent_tray_down=1

• 25: Operation home failed due to reagent_z not being able to move to reagent_tray_down=0 (System.ApplicationException)

• 26: Operation home failed due to reagent_z not being able to move to reagent_tray_down=0

• 30: Sensor align flow not aligned to 1

• 34: Operation home failed due to flowcell y not being able to move to flowcell_y_home=1

• 35: Sipper_Y not being able to move

• 36: Operation home failed due to sipper_y not being able to move to sipper_y_home=1 - fluidics

• 39: Cannot perform pierce. Machine state is unk and is expected to be loaded. The mechanisms have been automatically reinitialized in order to return to a working state

• 41: Cannot perform unload. Machine state is unk

• 47: Operation up failed due to straw_z not being able to move to straws_down=0

• 50: Operation up failed due to straw_z not being able to move to straws_up =1

• 51: Operation up failed due to straw_z not being able to move to straws down = 0

• 52: Operation down failed to waste presence not being 1

• 54: Operation down failed due to straw_z not being able to move to straws_down=1.

• 55: Operation up failed due to straw_z not being able to move to straws_down=0

• 58: Related to not loading tray

• 65: Sensor reagent_door_open should be 0, but it is not

• 66: Sipper_y stopped due to safety_interlock

• 67: Reagent_z stopped due to safety_interlock

• 69: Operation init failed due to flowcell_align1 not being 1 (or 0)

Troubleshooting steps:

1. First, reinsert the waste/spent reagent container (all the way to the back of the compartment).

2. Power cycle\* NextSeq 500/550.

• If control software fails to initialize with same notification, contact Illumina Technical Support for further guidance.

3. Perform a Quick Wash.

• If wash completes without error or all system check tests pass:

  • Proceed with the next run and monitor performance. See NextSeq 550 system specifications to verify performance.

• If the error reoccurs or poor run performance is observed, contact Illumina Technical Support for further guidance.

For reference, please see Knowledge Base articles:

• *How to perform a NextSeq 500/550 Power Cycle

• How to perform washes on the NextSeq 500/550

• How to perform a NextSeq 550 system check

For any feedback or questions regarding this article (Illumina Knowledge Article #1384), contact Illumina Technical Support [e-mail].