- Materials
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- Flow Cell Wash Kit (EXP-WSH003)
- Sequencing Auxiliary Vials (EXP-AUX001)
- Flow Cell Priming Kit (EXP-FLP002)
- Equipment
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- P1000 pipette and tips
- P20 pipette and tips
- Ice bucket with ice
- MinION Flow Cell
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To run a second library straight away, follow the instructions in the "Priming and loading the flow cell" section of your library preparation protocol with the recommendations below.
Note: For more consistent and better performance after a flushing a flow cell with the Flow Cell Wash Kit, we recommend the following:
- Pipette very slowly when loading priming mix into the flow cell.
- Wait five minutes between priming mix flushes.
- After the five minute pause, close the priming port, ensure the SpotON port is closed and remove the waste from waste port 1. This prevents the nuclease from diffusing through the flow cell. Repeat this step after the second priming mix flush.
As part of this process, the flow cell will need priming using flow cell priming reagents either in your sequencing kit or separately in the Flow Cell Priming Kit.
Once the flow cell has been primed and loaded, either resume the run in MinKNOW or start a new sequencing experiment. Use Join existing experiment when setting up a sequencing run to use the same settings that were used in a previous experiment.
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Reloading a library
Additional buffers are required for reloading a library following the washing of a flow cell. These can be found in the one of the following expansion kits:
Sequencing Auxiliary Vials V14 (EXP-AUX003). This expansion contains vials of Sequencing Buffer (SB), Elution Buffer (EB), Library Solution (LIS), and Library Beads (LIB) with Kit 14 flow cell priming reagents: Flow Cell Flush (FCF) and Flow Cell Tether (FCT). This expansion is only compatible with our Kit 14 chemistry e.g. SQK-LSK114.
Flow Cell Priming Kit (EXP-FLP004). This expansion contains both Kit 14 flow cell priming reagents required: Flow Cell Flush (FCF) and Flow Cell Tether (FCT). This expansion is only compatible with Kit 14 chemistry.
For our previous chemistries:
Sequencing Auxiliary Vials expansion (EXP-AUX001). This expansion contains vials of Elution Buffer (EB), Sequencing Buffer (SQB), and Loading Beads (LB), additional to those found in DNA sequencing kits for our Kit 9 chemistry.
Sequencing Auxiliary Vials expansion (EXP-AUX002). This expansion contains vials of Sequencing Buffer II (SBII), Elution Buffer (EB), Loading Solution (LS), and Loading Beads II (LBII), additional to those found in Kit 10 and 11 chemistry, such as:
- Kit 10 e.g. SQK-LSK110
- Kit 11 e.g. SQK-PCS111
- Flow Cell Priming Kit (EXP-FLO002). This expansion contains both flow cell priming reagents required: Flush Buffer (FB) and Flush Tether (FLT). This expansion is compatible with Kit 9, 10 and 11 chemistry.