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How to start sequencing
The sequencing device control, data acquisition, real-time basecalling, and barcode demultiplexing are carried out by the MinKNOW software. It is assumed you have already installed MinKNOW on your computer. Instructions for this can be found in the MinKNOW protocol.
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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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Select your connected device.
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Sequencing set-up
Select Start sequencing.
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Select positions
Enter your Experiment name and sample ID as prompted.
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Kit selection
The kit selection tab will provide a dropdown of available kits. Check the SQK-RBK004 kit.
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Run Options
The run options tab provides variables for run time and starting voltage. Leave these at the default values, except the run time which can be set to 12 hours.
Active Channel Selection refers to a feature introduced in MinKNOW v2.2 If a channel is in the "Saturated" or "Multiple" state, the software instantly switches to a new channel in the group. If a channel is "Recovering", MinKNOW will attempt to revert the channel back to "Pore" or "Sequencing" for ~5 minutes, after which it will select a new channel in the group. This maximises the number of channels sequencing at the start of the experiment.
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Basecalling
Check Basecalling and select the Fast basecalling option in the dropdown menu.
Select Barcoding to Enabled. -
Output
Select where you would like to save the data.
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Review
Review your sequencing set-up and click Run.