ONT Flow Cell Wash Kit XL

The Flow Cell Wash Kit is a valuable accessory for maintaining and maximizing the performance of Oxford Nanopore flow cells. It enables users to wash and reuse flow cells, extending their utility for multiple sequencing runs. The kit effectively removes loaded libraries, reverts "unavailable" pores to the "single pore" state, and minimizes contamination between runs.

Key Features

1. Flow Cell Reusability:
   • Allows users to remove existing libraries from flow cells, enabling their reuse for fresh samples.
   • Supports efficient sample-on-demand workflows without batching.
2. Enhances Pore Availability:
   • Reverts pores from the "unavailable" state to the "single pore" state, increasing the number of active pores.
   • Proven to significantly boost flow cell output, with output more than doubling in some cases after repeated washes.
3. Minimal Contamination:
   • Utilizes a nuclease-based cleaning process with contamination levels as low as 0.1%.
   • Enables accurate barcoded filtering in cases where minor contamination may persist.
4. Nuclease Inactivation:
   • After washing, the nuclease is inactivated to prevent residual activity from affecting subsequent runs.
5. Maximizing Output Over Time:
   • Multiple wash steps can significantly prolong the life of a flow cell by maintaining active pores and consistent data acquisition rates.

Kit Contents

Applications

1. Sequential Runs:
   • Ideal for users running different samples sequentially without needing new flow cells.
2. On-Demand Sample Processing:
   • Enables non-batched workflows for flexibility in sequencing experiments.
3. Reactivation of Unavailable Pores:
   • Addresses the reduction in data acquisition rates caused by "recovering" or "unavailable" pores.
4. Extending Flow Cell Lifespan:
   • Supports cost-effective sequencing by reusing flow cells for additional libraries.

Workflow

1. Library Removal:
   • Use the Wash Mix to digest and remove loaded nucleic acids.
   • Perform a wash to ensure efficient removal of all traces of the library.
2. Nuclease Inactivation:
   • Following the wash step, the nuclease is inactivated to prepare the flow cell for reuse.
3. Reuse or Storage:
   • Reload the flow cell with a new library immediately or store it using the Storage Buffer for later use.
4. Monitoring Performance:
   • Use MinKNOW to monitor the number of active pores before and after the wash procedure.

Performance Results

• Increased Pore Availability:
  • Flow cells washed with the kit saw significant recovery in the number of single pores, enabling higher data acquisition rates.
• Minimal Residual Activity:
  • Subsequent runs show no deterioration in read lengths, indicating effective nuclease inactivation.
• Doubling of Output:
  • In PromethION runs, sequential washes doubled output by maintaining pore availability.

Compatibility

Kits:

• Compatible with all ONT sequencing kits.

Flow Cells:

• FLO-MIN106D
• FLO-MIN114
• FLO-PRO002
• FLO-PRO114M

Devices:

• MinION Mk1B
• MinION Mk1C
• GridION
• PromethION

Shipping and Storage

• Shipping Conditions:
  • Shipped at 2–8°C in temperature-controlled packaging.
• Storage Conditions:
  • Store at -20°C for long-term stability.
• Shelf Life:
  • Product deemed usable for 3 months from receipt.

Advantages

1. Cost Efficiency:
   • Extends the lifespan of expensive flow cells, reducing costs associated with frequent replacements.
2. Optimized Output:
   • Maintains high data acquisition rates by reactivating pores.
3. Flexibility:
   • Allows reloading and reusing flow cells for on-demand sequencing.
4. Sustainability:
   • Reduces waste by enabling the reuse of flow cells.

Additional Notes

• Barcoded Filtering:
  • For sequential runs with minor contamination risks, barcoding ensures accurate filtering of previous samples.
• Additional Reagents:
  • Reloading washed flow cells requires reagents from the Sequencing Auxiliary Vials Expansion (EXP-AUX001 or EXP-AUX110).

The Flow Cell Wash Kit is an essential tool for labs aiming to maximize the utility of ONT flow cells, enabling efficient sequencing workflows, cost savings, and high-quality data output over multiple uses.