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 errors 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. Re-insert the waste/spent reagent container (all the way to the back of the compartment).

2. Power cycle the NextSeq 500/550. For detailed instructions, see How to Power Cycle the NextSeq 500/550.

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

1. Perform a Quick Wash. For detailed instructions, see How to perform washes on the NextSeq 500/550.

2. If the wash completes normally, perform a system check. For detailed instructions, see How to perform a NextSeq 500/550 system check.

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