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Calculating pore occupancy
Simplex data is generated when a single strand of DNA is sequenced. It happens at a much higher volume than duplex sequencing, meaning higher outputs can be generated.
For experiments that require high data output, we recommend loading your flow cell with a high amount of library (35-50 fmol) to ensure high pore occupancy of >95% is reached to maximise data output. If your sequencing run starts with a lower pore occupancy after 1 hour of starting, data output may be negatively impacted.
Pore occupancy is the percentage of pores that are sequencing at any time out of those available for sequencing. This is displayed in the pore activity plot in MinKNOW during an experiment. During the first few minutes of sequencing, pore occupancy can increase as the library tethers to the membrane. Therefore, we recommend reviewing pore occupancy ~1 hour into a sequencing run.
To calculate pore occupancy: