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How to start sequencing
The sequencing device control, data acquisition and real-time basecalling are carried out by the MinKNOW software. It is assumed you have already installed MinKNOW on your computer. Further instructions for setting up a sequencing run can be found in the MinKNOW protocol.
We recommend setting up a sequencing run on a MinION or GridION device using the basecalling and barcoding recommendations outlined below. All other parameters can be left to their default settings.
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Open the MinKNOW software using the desktop shortcut and log into the MinKNOW software using your Community credentials.
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Click on your connected device.
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Set up a sequencing run by clicking Start sequencing.
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Select the Rapid Barcoding Kit 24 V14 (SQK-RBK114.24) or Rapid Barcoding Kit 96 V14 (SQK-RBK114.96).
Click Continue to Run Options to continue.
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Change the run limit to 12 hours by clicking "Options" and changing the run limit value to 12. The other run settings can be left at the defaults.
Click Continue to basecalling to continue.
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Set up basecalling and barcoding using the following parameters:
Ensure basecalling is ON.
Next to "Models", click Edit options and choose High accuracy basecaller (HAC) from the drop-down menu.
Ensure barcoding in ON.
All other options can be kept to their default settings.
Click Continue to output and continue.
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Set up the output format and filtering as follows:
Select either .POD5 or .FAST5 (legacy) as the output format.
Ensure .FASTQ is selected for basecalled reads.
Ensure filtering is ON and read splitting is enabled. Other parameters can be kept to their default settings.
Click Continue to final review to continue.
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Click "Start" to start sequencing.
You will be automatically navigated to the "Sequencing Overview" page to monitor the sequencing run.