- Materials
-
- Flow Cell Wash Kit XL (EXP-WSH004-XL)
- Flow cell priming reagents available in your sequencing kit or in the following kits:
- Sequencing Auxiliary Vials V14 (EXP-AUX003)
- Flow Cell Priming Kit V14 (EXP-FLP004)
- Consumables
-
- PromethION Flow Cell
- 1.5 ml Eppendorf DNA LoBind tubes
- 2 ml Eppendorf DNA LoBind tubes
- Equipment
-
- P1000 pipette and tips
- P200 pipette and tips
- P20 pipette and tips
- Ice bucket with ice
-
To run a second library straight away, follow the instructions in the 'Priming and loading multiple flow cells on a PromethION' of this 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.
The flow cells will already be inserted in the device and will not need a second flow cell check but will require priming with flow cell priming reagents. Once all the flow cells have been primed, either resume your run in MinKNOW or start a new sequencing experiment using Join existing experiment to use the same settings that were used previously.
-
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.