-
Overview of nanopore data analysis
For a full overview of nanopore data analysis, which includes options for basecalling and post-basecalling analysis, please refer to the Data Analysis document.
-
How to start sequencing
The sequencing device control, data acquisition and real-time basecalling are carried out by the MinKNOW software. You will need to have MinKNOW already installed on your computer before starting.
Data acquisition and basecalling in real-time using MinKNOW on a computer
For references, please refer to the MinKNOW protocol.
Instructions on how to carry out sequencing:
- Double-click the MinKNOW icon on the desktop to open the MinKNOW GUI and log in with your nanopore community credentials
- With the MinION connected to the computer select the sequencing device connected
- On the Start homepage, select 'Start Sequencing' and complete the run parameters for the experiment
- Provide an experiment name: we recommend using the date for the run
- Provide a Sample ID for the run
- Select the type of flow cell being used from the drop-down menu and continue to kit selection
- Select the kit used to prepare the libraries: SQK-RPB004
- Set the run time to 48 hours. However, samples positive for mpox will likely show a positive result after ~8 hours
- Select 'Continue to Basecalling'
- Select 'High accuracy basecalling' (HAC)
- Check 'Barcoding' on
- Select the output data location, format and filtering options. Output data is typically saved as FAST5 and/or FASTQ
- Continue to 'Run Setup'
- Start run
- Navigate to 'Sequencing Overview' to monitor the run