- Materials
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- DNA libraries
- Flow Cell Priming Kit V14 (EXP-FLP004)
- Sequencing Auxiliary Vials V14 (EXP-AUX003)
- Flow Cell Wash Kit XL (EXP-WSH004-XL)
- Consumables
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- PromethION Flow Cell (FLO-PRO114M)
- 1.5 ml Eppendorf DNA LoBind tubes
- 2 ml Eppendorf DNA LoBind tubes
- Equipment
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- PromethION sequencing device
- P1000 pipette and tips
- P200 pipette and tips
- P20 pipette and tips
- Ice bucket with ice
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This protocol has been written for Kit 14 chemistry but can be used for all our library preparation protocols.
We recommend using this as a guideline for how to prepare multiple flow cells alongside the relevant protocol for your sequencing kit.
To wash your flow cells, the Flow Cell Wash Kit XL (EXP-WSH004-XL) is required.
To reload the flow cell with a DNA library, the following additional reagents are required.
- Flow Cell Tether (FCT)
- Flow Cell Flush (FCF)
- Library Beads (LIB) or Library Solution (LIS)
- Sequencing Buffer (SB)
- Elution Buffer (EB)
These can all be found in the Sequencing Auxiliary Vials V14 (EXP-AUX003). If only flow cell priming reagents are required, these can be found in the Flow Cell Priming Kit V14 (EXP-FLP004).
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Flow Cell Wash Kit XL (EXP-WSH004-XL) contents
Name Acronym Fill volume per vial (µl) Cap colour No. of vials No. of uses Wash Mix WMX 150 Brown 1 48 Wash Diluent DIL 20,000 White cap, brown stripe 1 48 Storage Buffer S 25,000 White cap, yellow stripe 1 48 - Wash Mix (WMX) contains DNase I.
- Wash Diluent (DIL) contains the exonuclease buffer that maximises activity of the DNase I.
- The Storage Buffer allows flow cells to be stored for extended periods of time.
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Sequencing Auxiliary Vials V14 (EXP-AUX003) contents
Name Acronym Cap colour No. of vials Fill volume per vial (μl) Elution Buffer EB Black 2 500 Sequencing Buffer SB Red 2 700 Library Solution LIS White cap, pink label 2 600 Library Beads LIB Pink 2 600 Flow Cell Flush FCF Light blue label 2 8,000 Flow Cell Tether FCT Purple 2 200 -
Flow Cell Priming Kit (EXP-FLP004) contents
Name Acronym Cap colour No. of vials Fill volume per vial (μl) Flow Cell Flush FCF 6 Clear cap, light blue lable 8,000 Flow Cell Tether FCT 1 Purple 200