A flow cell used in Nanopore sequencing can typically last for up to 72 hours in a single, continuous sequencing run. This duration represents the maximum operational time for a single experiment to maximize data collection from the same sample.
While a single run can extend up to 72 hours, the overall utility and reusability of a flow cell can vary depending on several factors, including the initial number of active pores, sample type, and experimental conditions.
Factors Influencing Flow Cell Lifespan and Performance
The longevity and efficiency of a flow cell are influenced by a combination of inherent characteristics and operational practices. Understanding these factors is crucial for optimizing sequencing experiments.
Key Factors Affecting Flow Cell Lifespan
| Factor | Description | Impact on Lifespan |
|---|---|---|
| Active Pore Count | The number of functional nanopores available for sequencing at the start of a run. | Directly correlates with data output and potential for reusability. Drops over time due to pore blockages or damage. |
| Sample Quality | Purity, concentration, and integrity of the nucleic acid sample. Contaminants or highly degraded samples can clog or damage pores. | Poor quality samples can rapidly reduce active pores, shortening effective lifespan. |
| Run Conditions | Temperature, buffer stability, and the presence of inhibitory substances in the library. | Optimal conditions maintain pore activity; harsh conditions can accelerate degradation. |
| Washing & Storage | Proper cleaning and storage procedures between runs or prior to a run. | Incorrect handling or storage can lead to pore degradation or contamination, reducing lifespan. |
| Application Needs | Whether maximizing yield from a single sample or performing multiple, shorter runs with different samples. | Dictates how a flow cell is used; shorter runs might allow for more reuses. |
Maximizing Flow Cell Performance and Longevity
To get the most out of your flow cell and potentially extend its effective lifespan, consider the following best practices:
- Optimal Sample Preparation: Ensure your DNA or RNA library is clean, pure, and free from contaminants that could clog or inactivate nanopores. High-quality input is paramount for sustained sequencing.
- Proper Priming and Loading: Follow manufacturer guidelines meticulously for priming the flow cell and loading your library to ensure even distribution and minimize air bubbles.
- Post-Run Washing: If planning to reuse a flow cell, perform a thorough washing procedure immediately after a run. This helps remove residual DNA, proteins, and contaminants, preserving pore health.
- Careful Storage: Store flow cells according to manufacturer specifications (e.g., specific temperatures, upright position) to maintain their integrity over time.
- Monitor Pore Activity: Utilize the real-time monitoring features of Nanopore sequencing software to track the number of active pores. This allows for timely decisions on when to stop a run or if a flow cell is suitable for another experiment.
Flow Cell Reusability
While a single continuous run can last up to 72 hours, flow cells can sometimes be reused for subsequent sequencing runs if a sufficient number of active pores remain after the initial experiment and proper washing protocols are followed. The possibility of reuse extends the overall utility of the flow cell beyond a single run's duration, making it a cost-effective option for certain experimental designs. However, each subsequent run might yield less data due to a gradual decrease in active pores.
