How to perform a Maintenance Wash on the NovaSeq X/X Plus

The NovaSeq X/X Plus will perform an automated wash after every successful sequencing run. As part of normal instrument maintenance, an additional maintenance wash will be required every 14 days or when an automatic post-run wash fails to complete.

Note: For most sequencing run issues, the instrument will attempt to perform the automatic post-run wash so a maintenance wash will not be required for many run-stopping issues.

For detailed instructions, see the Perform a Maintenance Wash section of the NovaSeq X Plus Product Documentation.

Wash Buffer Preparation

1. Add 400 ml laboratory-grade water to a 500 ml centrifuge bottle.

2. Add 0.2 ml 100% Tween-20 to the laboratory-grade water to prepare the 0.05% Tween-20 wash solution.

• Using a freshly prepared dilution of Tween-20 limits the introduction of contaminants into the fluidics system.

3. Invert solution several times to mix thoroughly.

4. Add 0.05% Tween-20 wash solution to the wash solution well on the wash reagent cartridge.

5. Combine the following volumes in a 50 ml centrifuge tube to prepare 49 ml of 0.12% reagent grade NaOCl:

   1. 1.2 ml 5% reagent grade NaOCl.

   2. 48 ml Deionized (DI) water.

6. Invert solution several times to mix thoroughly.

7. Add 49 ml 0.12% NaOCl solution to the bleach well on the wash reagent cartridge.

Setting up the Maintenance Wash (Run Time: ~3 hours)

1. Select the instrument icon to open the global navigation menu.

2. Select Settings, and then select Wash.

3. Select the instrument side(s) to perform the wash on.

4. On the Wash screen, select Unload wash flow cells.

• After selecting, the display monitor raises and the flow cell door opens. The flow cell light indicates the instrument side that on which the wash is being performed.

5. If a used 8-lane flow cell is already loaded into the system, select Load wash flow cells.

6. If a used flow cell is not already loaded, load the wash flow cells onto the flow cell holder and select Load wash flow cells.

7. On the Wash screen, select Unload wash reagents.

• The instrument doors unlock automatically and the system displays information on loading the wash reagent and buffer cartridges.

8. Remove the used sequencing reagent cartridge, then load the wash reagent cartridge into the instrument in the right bay of the reagent drawer.

• The used buffer cartridge can be left in place.

9. Select Load wash reagents.

10. Empty the used reagent bottles then replace the used reagent bottles into the used reagent drawers.

• Dispose of spent reagents according to local EH&S guidelines.

11. Select Confirm after emptying and replacing the used reagent bottles.

12. Close the reagent drawers, then close the instrument doors.

13. After all wash consumables are loaded, select Start wash.

14. After starting the wash, the instrument doors automatically lock. The doors unlock after the wash completes and the instrument returns to the Start screen.

For any feedback or questions regarding this article (Illumina Knowledge Article #7479), contact Illumina Technical Support [e-mail].