ONT Flow Cell Wash (EXP-WSH004)
Nuclease wash of an Oxford Nanopore flow cell between sequencing runs using the Flow Cell Wash Kit (EXP-WSH004). Removes up to 99.9% of library DNA and adapters, allowing the flow cell to be reused for a subsequent library. Based on ONT EXP-WSH004 protocol.
Version History
Version 1.0 Current
Effective: 2026-02-23Procedure Details
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MinKNOW (ONT sequencing software)
Flow cell wash via MinKNOW UI -
Micropipette
Liquid handling
Micropipettes (P1000, P200) -
Vortex mixer
Mixing
Vortex mixer for wash preparation -
Nanopore sequencer
Sequencing
MinION / GridION / PromethION sequencer
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Flow Cell Wash Kit
(Oxford Nanopore)
EXP-WSH004Sequencing kit
Flow Cell Wash Kit (EXP-WSH004)
Procedure Steps (Version 1.0)
Ensure sequencing has completed and the run has been stopped in MinKNOW. Do not manipulate the flow cell while sequencing is active.
Thaw one tube of Flow Cell Wash Solution (WSH) at room temperature. Mix by inversion. Do not vortex. Place on ice once thawed.
Prepare Flow Cell Wash Mix: add 380 µL WSH to 20 µL Flow Cell Wash Diluent (DIL). Mix by pipetting. Place on ice. Use within 1 day of preparation.
Open the SpotON port and the priming port on the flow cell.
Draw back a small volume (20–30 µL) through the priming port using a P1000 pipette to ensure no air is trapped.
Load 200 µL of Flow Cell Wash Mix through the priming port. Avoid introducing air bubbles.
Wait 5 minutes to allow nuclease digestion of the library.
Load the remaining 200 µL of Flow Cell Wash Mix through the priming port.
Close the SpotON port and the priming port. Wait at least 5 minutes before re-priming and loading a new library.
Perform a flow cell check in MinKNOW to assess available pores before loading the next library. Record the pore count.
If the flow cell will not be used immediately, load Storage Buffer (S) from the wash kit and store at 2–8 °C.
References
- Oxford Nanopore Technologies — Flow Cell Wash Kit (EXP-WSH004) Protocol, Rev B, 26 Sep 2024 Link protocol