- Materials
-
- 200 ng gDNA per sample
- Rapid Barcoding Kit 24 V14 (SQK-RBK114.24) OR Rapid Barcoding Kit 96 V14 (SQK-RBK114.96)
- Consumables
-
- MinION and GridION Flow Cell
- Qubit dsDNA HS Assay Kit (Invitrogen, Q32851)
- Bovine Serum Albumin (BSA) (50 mg/ml) (e.g Invitrogen™ UltraPure™ BSA 50 mg/ml, AM2616)
- Freshly prepared 80% ethanol in nuclease-free water
- Nuclease-free water (e.g. ThermoFisher, AM9937)
- Qubit™ Assay Tubes (Invitrogen, Q32856)
- Eppendorf twin.tec® PCR plate 96 LoBind, semi-skirted (Eppendorf™, cat # 0030129504) with heat seals
- 2 ml Eppendorf DNA LoBind tubes
- 0.2 ml thin-walled PCR tubes
- 1.5 ml Eppendorf DNA LoBind tubes
- Equipment
-
- MinION or GridION device
- MinION and GridION Flow Cell Light Shield
- Microplate centrifuge, e.g. Fisherbrand™ Mini Plate Spinner Centrifuge (Fisher Scientific, 11766427)
- Timer
- Thermal cycler or heat blocks
- Magnetic rack
- Hula mixer (gentle rotator mixer)
- Ice bucket with ice
- Qubit fluorometer (or equivalent for QC check)
- P1000 pipette and tips
- P200 pipette and tips
- P100 pipette and tips
- P20 pipette and tips
- P2 pipette and tips
- Multichannel pipette and tips
-
Rapid Barcode use requirements
Note: Ensure you are using the correct kit for your desired number of samples:
- Rapid Barcoding Kit 24 V14 (SQK-RBK114.24) contains barcodes RB01-24, allowing you to multiplex up to 24 samples.
- Rapid Barcoding Kit 96 V14 (SQK-RBK114.96) contains barcodes RB01-96, allowing you to multiplex up to 96 samples.
For optimal output, we currently do not recommend using fewer than 4 barcodes. If you wish to multiplex less than 4 samples, please ensure you split your sample(s) across multiple barcodes so at least 4 barcodes are run (e.g. for 2 samples, use RB01-RB02 for sample A and RB03-RB04 for sample B).
Please note that the required sample input for each barcode is 200 ng gDNA.Alternatively, you might want to explore our Rapid Sequencing Kit V14 (SQK-RAD114) for sequencing individual or small sets of samples.
-
Check your 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/GridION/PromethION or within four weeks of purchasing Flongle Flow Cells. Oxford Nanopore Technologies will replace any flow cell with fewer than the number of pores in the table below, 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.
Flow cell Minimum number of active pores covered by warranty Flongle Flow Cell 50 MinION/GridION Flow Cell 800 PromethION Flow Cell 5000 -
Rapid Barcoding Kit 24 V14 (SQK-RBK114.24) contents
Name Acronym Cap colour No. of vials Fill volume per vial (µl) Rapid Adapter RA Green 1 15 Adapter Buffer ADB Clear 1 100 AMPure XP Beads AXP Amber 2 1,200 Elution Buffer EB Black 1 500 Sequencing Buffer SB Red 1 700 Library Beads LIB Pink 1 600 Library Solution LIS White cap, pink label 1 600 Flow Cell Flush FCF Blue 6 1,170 Flow Cell Tether FCT Purple 1 200 Rapid Barcodes RB01-24 Clear 24 15 This Product Contains AMPure XP Reagent Manufactured by Beckman Coulter, Inc. and can be stored at -20°C with the kit without detriment to reagent stability.
-
Rapid Barcoding Kit 96 V14 (SQK-RBK114.96) contents
Name Acronym Cap colour No. of vials Fill volume per vial (µl) Rapid Adapter RA Green 2 15 Adapter Buffer ADB Clear 1 100 AMPure XP Beads AXP Amber 3 1,200 Elution Buffer EB Black 1 1,500 Sequencing Buffer SB Red 1 1,700 Library Beads LIB Pink 1 1,800 Library Solution LIS White cap, pink label 1 1,800 Flow Cell Flush FCF Clear 1 15,500 Flow Cell Tether FCT Purple 2 200 Rapid Barcodes RB01-96 - 3 plates 8 µl per well This Product Contains AMPure XP Reagent Manufactured by Beckman Coulter, Inc. and can be stored at -20°C with the kit without detriment to reagent stability.