- Materials
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- Flow Cell Priming Kit (EXP-FLP004)
- Sequencing Auxiliary Vials V14 (EXP-AUX003)
- Flow Cell Wash Kit (EXP-WSH004) or Flow Cell Wash Kit XL (EXP-WSH004-XL)
- SFB Expansion (EXP-SFB001)
- Consumables
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- MinION Flow Cell (FLO-MIN106 or FLO-MIN114)
- Bovine Serum Albumin (BSA) (50 mg/ml) (e.g Invitrogen™ UltraPure™ BSA 50 mg/ml, AM2616)
- 1.5 ml Eppendorf DNA LoBind tubes
- Agencourt AMPure XP beads (Beckman Coulter, A63881)
- Qubit™ Assay Tubes (Invitrogen, Q32856)
- Qubit dsDNA HS Assay Kit (Invitrogen, Q32851)
- Equipment
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- MinION or GridION device
- P1000 pipette and tips
- P200 pipette and tips
- P20 pipette and tips
- Ice bucket with ice
- Vortex mixer
- Hula mixer (gentle rotator mixer)
- Microfuge
- Magnetic rack
- Heating block
- Qubit fluorometer (or equivalent)
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Check your MinION flow cell
We highly recommend that you check the number of pores in your flow cell prior to starting a sequencing experiment. This should be done within three months of purchasing for MinION flow cells. Oxford Nanopore Technologies will replace any flow cell with fewer than 800 pores when the result is reported within two days of performing the flow cell check, and when the storage recommendations have been followed. To do the flow cell check, please follow the instructions in the Flow Cell Check document.
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Flow cell priming reagents
Ensure the compatible flow cell priming reagents are used for your libraries. These are required for all methods and are available with our sequencing kits. If extra reagents are required, they can be found in the following expansion kits.
For libraries prepared using Kit 14 chemistry, please ensure you are using the following reagents:
- Flow Cell Tether (FCT)
- Flow Cell Flush (FCF)
- For MinION flow cells only, we recommend using Bovine Serum Albumin (BSA)
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 For libraries prepared using Kit 9, 10 or 11 chemistry, please ensure you are using the following reagents:
- Flush Buffer (FB)
- Flush Tether (FLT)
Flow Cell Priming Kit (EXP-FLP002) contents:
Name Acronym Cap colour No. of vial Fill volume per vial (μl) Flush Buffer FB Blue 6 1,170 Flush Tether FLT Purple 1 200 -
Short Fragment Buffer (SFB) and Elution Buffer (EB)
Short Fragment Buffer (SFB) and Elution Buffer (EB) are both required for the "Clean up and transfer a library between flow cells" method. Both reagents are found in our sequencing kits but extra reagents can be found in the below kits.
Short Fragment Buffer (SFB) is available in the SFB Expansion (EXP-SFB001).
SFB Expansion (EXP-SFB001) contents:
Name Acronym Cap colour No. of vials Fill volume per vial (μl) Short Fragment Buffer SFB Grey 4 1,800
Elution Buffer (EB) is available in the Sequencing Auxiliary Kit.For libraries prepared using Kit 14 chemistry, please ensure you are using the following kit which also includes the flow cell priming reagents:
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 For libraries prepared using the Kit 10 or 11 chemistry, please use the following kit:
Sequencing Auxiliary Vials (EXP-AUX002) contents:
Name Acronym Cap colour No. of vials Fill volume per vial (μl) Sequencing Buffer SBII Red 4 500 Loading Solution LS Pink sticker on cap 2 360 Elution Buffer EB Black 2 200 Loading Beads LBII Pink 2 360 For libraries prepared using Kit 9 chemistry, please use the following kit:
Sequencing Auxiliary Vials (EXP-AUX001) contents:
Name Acronym Cap colour No. of vials Fill volume per vial (μl) Sequencing Buffer SQB Red 6 300 Elution Buffer EB Black 2 200 Loading Beads LB Pink 2 360 -
AMPure XP Beads (AXP)
For the "Clean up and transfer a library between flow cells" method, AMPure XP Beads (AXP) are required. These are available in many of our kits where the DNA library undergoes a clean up step. However, if extra beads are required, we recommend purchasing more from Beckman Coulter, Inc. (A63880).
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Flow Cell Wash Kit
The Flow Cell Wash Kit is required for the "Recover a library to replace on a washed flow cell" method.
Flow Cell Wash Kit contents (EXP-WSH004):
Contents Volume (µl) No. of tubes No. of uses Wash Mix (WMX) 15 1 6 Wash Diluent (DIL) 1,300 2 6 Storage Buffer (S) 1,600 2 6