Wash best practices for the MiSeq
Many factors contribute to a successful sequencing run. One important aspect is to make sure that the fluidics lines are thoroughly flushed between sequencing runs to prevent any precipitate accumulation. It is also important that the fluidics systems are well maintained when the instrument is idle.
This article outlines all different types of washes on the MiSeq and when they should be performed.
Note: When any wash is complete, leave the used flow cell, wash tray, and wash bottle containing the remaining wash solution in the instrument. The sippers remain in the down position, preventing the sippers from drying out and air from entering the system.
Times for washes:
Post-Run wash: 20 minutes.
Post-Run wash with Template line: 30 minutes.
Manual or Maintenance wash: 90 minutes.
Standby wash: 2 hours.
Post-Run Wash:
Required after every sequencing run.
One wash step with 0.5% Tween 20 wash solution.
Add 6 ml wash solution to each reservoir of the wash tray.
Add 350 ml wash solution to the 500 ml wash bottle.
Wash volume expected in the waste bottle: 17.25 ml.
Template Line Wash:
Optional for Research Use Only (RUO) runs.
Additional step to regular post-run wash, with a 0.5% Tween 20 wash solution.
Add 6 ml wash solution to each reservoir of the wash tray.
Add 350 ml wash solution to the 500 ml wash bottle.
Add 0.01% sodium hypochlorite (NaOCl, lab grade bleach) to the disposable wash tube in position 17 of the wash tray.
Wash volume expected in the waste bottle: 25.5 ml.
Maintenance Wash:
Required every 30 days.
The instrument will display a warning notification that a maintenance wash is required 30 days from the last maintenance wash.
The instrument will prevent the user from starting a new sequencing run if a maintenance wash has not been completed within the previous 45 days.
Includes three wash steps with 0.5% Tween 20 wash solution (fresh for each wash step).
Add 6 ml wash solution to each reservoir of the wash tray.
Add 350 ml wash solution to the 500 ml wash bottle.
Wash volume expected in the waste bottle: 51.75 ml.
Standby Wash:
Prepares instrument for idle mode.
Perform a standby wash if the instrument is unused for > 7 days and every 30 days thereafter if the instrument remains idle.
Includes two wash steps with 0.5% Tween 20 wash solution (fresh for each wash step).
Add 6 ml wash solution to each reservoir of the wash tray.
Add 350 ml wash solution to the 500 ml wash bottle.
When the standby wash is complete, the instrument will be in standby mode and a message appears on the home screen indicating the status of the instrument.
To return the instrument to use, power cycle the instrument to reset communications and complete a maintenance wash to bring the instrument out of standby mode.
Wash volume expected in the waste bottle: 46 ml.
For any feedback or questions regarding this article (Illumina Knowledge Article #1679), contact Illumina Technical Support techsupport@illumina.com.
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