# 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:

![](https://761066130-files.gitbook.io/~/files/v0/b/gitbook-x-prod.appspot.com/o/spaces%2FGM9W2DuBTgEXv1ClCm8H%2Fuploads%2Fgit-blob-7baf37a8dc30bae998e64517c5f5a5066102ef1d%2Fimage1.jpg?alt=media)

**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](https://www.illumina.com/company/contact-us.html#/united-states/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](https://www.illumina.com/systems/sequencing-platforms/nextseq/specifications.html) to verify performance.
* If the error reoccurs **or** poor run performance is observed, contact [Illumina Technical Support](https://www.illumina.com/company/contact-us.html#/united-states/technical-support) for further guidance.

For reference, please see Knowledge Base articles:

* **\***[**How to perform a NextSeq 500/550 Power Cycle**](https://knowledge.illumina.com/instrumentation/nextseq-500-550/instrumentation-nextseq-500-550-reference_material-list/000001972)
* [**How to perform washes on the NextSeq 500/550**](https://knowledge.illumina.com/instrumentation/nextseq-500-550/instrumentation-nextseq-500-550-reference_material-list/000003076)
* [**How to perform a NextSeq 550 system check**](https://knowledge.illumina.com/instrumentation/nextseq-500-550/instrumentation-nextseq-500-550-troubleshooting-list/000001883)

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