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The PromethION 24 and 48
This is a benchtop device designed to run up to 24 (for P24) or 48 (for P48) flow cells. Each flow cell is independently addressable, meaning that experiments can be run concurrently or individually. The integrated compute in the PromethION enables real-time data streaming and analysis. The device is operated by the MinKNOW software that controls the device, experimental scripts, and performs basecalling.
Below is a quick start guide to set up a new device and to start sequencing your library samples. For further details using this device, please see the PromethION 24/48 User Manual.
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Install the PromethION device.
Unpack the device and place the Sequencing Unit and the Data Acquisition Unit within 2 meters of each other.
Connect the following cables:
- Connect either the copper or fibre cables to the data acquisition unit.
- Connect the PCIe and USB cables between the sequencing and data acquisition units
- Connect the peripheral devices (e.g. monitor, mouse, keyboard) to the appropriate ports.
- Connect the two C13 cables to both units.
Turn on the power at the mains supply and turn on the sequencing unit, wait three minutes and turn on the data acquisition unit.
Validate your IP address to ensure it has been correctly set using a static or DHCP address using the command
ifconfig
.
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Log into MinKNOW.
Double-click on the MinKNOW icon on your desktop.
Click on Log in with your Nanopore account.
Enter your email address and Nanopore password.
The Connection Manager will open, and your device will be the card in the top left corner.
Click on My device card to access and control it.
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Tutorials
These are available to help you get started and run your experiments as easily as possible.
On opening the MinKNOW software for the first time, the tutorials will navigate you through the graphical user interface (GUI). Click Continue tutorial to follow or click the ⁞ to skip.
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Check the time and date of your device.
Verify the time and data of your device is correct to successfully establish a network connection.
Navigate to Device settings in the host settings.
Check the time and date.
Click Change data and time and set the time using custom date/time.
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Reset the default password.
The device is shipped with a default password
prom
that must be reset.Navigate to Device settings in the host settings.
Click Reset password to open the Change password dialogue box.
Enter the current password, followed by the new password.
Click Reset password to confirm the password change.
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Perform a hardware check.
A hardware check must be performed on new devices or after a software update.
Open the PromethION and insert the CTCs into all positions. The LED adjacent to the flow cell will be blue when initialising and will turn green when it is ready.
Close the device lid.
Navigate to the Start page and click Hardware check.
Click Select all available positions.
Click Start and the hardware check will complete after approximately 1 minute.
After the hardware check has finished, remove the CTCs and store in dry conditions.
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Perform a flow cell check.
A flow cell check must be carried out before loading a DNA or RNA library to assess the number of pores available.
Open the device and insert your flow cell.
Navigate to the Start page and click Flow Cell Check.
MinKNOW will recognise the PromethION Flow Cell type and ID.
Click Start and the flow cell check will begin.
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Set up a sequencing run.
Prime and load your flow cell with DNA or RNA library by following a library prep protocol and set-up a sequencing run, as follows:
Navigate to the Start page and click Start Sequencing.
In the Positions tab, fill in the experiment name, sample ID, and select the flow cell type and position(s). Click Continue to kit selection.
Select the kit and any expansion(s) used to prepare the library. Click Continue to run options.
Specify the sequencing run length and minimum read length or keep the default settings. Click Continue to analysis.
Choose your basecaller model and select any available barcoding and alignment options or keep the default settings. Click Continue to output.
Specify your output data location, format and filtering options or keep the default settings. Click Continue to final review.
Click Start.
For further details, please see the Starting a sequencing run section for your device in the MinKNOW protocol.