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London Calling 2024
  21st - 24th May
16S Barcoding Kit 24 V14
SQK-16S114.24

Genus-level bacterial identification with barcoding for up to 24 samples.

Product lead time: 1 week

This uses our latest Kit 14 chemistry.

£810.00
3Released
  • Products from Oxford Nanopore Technologies that have completed their Early Access
  • Users can continue to provide feedback through feature request pinboard
  • Warranty periods extended to 1 – 3 months
  • Product continues to iterate with upgrade notifications of up to 3 months
  • Lead time visible in store
  • Product subject to availability

Information

This kit is recommended for users who:

  • want to do 16S sequencing
  • are interested in genus-level bacterial identification

Please note that to use this kit, you will need to purchase additional 3rd party reagents: see the "3rd Party Materials" tab for more detail.

Multiplexing samples onto one flow cell can reduce the cost per sample for a user. Below are some worked examples:

No barcodes12 barcodes24 barcodes24 barcodes (Flongle)
Flow cell price$500$500$500$90 (Flongle flow cell)
Library price$90$120$150$150
Price per sample$590$51.67$27.08$10

The 16S Barcoding Kit 24 V14 offers a method of amplifying the entire ~1,500 bp 16S rRNA gene from extracted gDNA, followed by library preparation. By narrowing down to a specific region of interest, a user can see all the organisms in the sample without sequencing unnecessary regions of the genome, making the identification quicker and more economical. On the other hand, analysing the full 16s rRNA gene is more informative than analysing a subset of exons.

The 16S Barcoding Kit 24 V14 features:

FeatureProperty
Preparation time25 minutes + PCR
Input requirement10 ng gDNA per sample
PCR RequiredYes
FragmentationNot required
Read lengthFull-length 16S gene (~1.5 kb)
Kit chemistryKit 14
Associated protocolsRapid sequencing DNA - 16S Barcoding Kit 24 V14 (SQK-16S114.24)
Multiplexing options• 16S Barcoding Kit 24 V14
• Flow Cell Wash Kit
Pack size6 reactions
StabilityShipped at 2–8°C

Long-term storage -20°C

Oxford Nanopore Technologies deem the useful life of the product to be 3 months from receipt by the customer

The DNA is amplified by PCR using specific 16S primers (27F and 1492R) that contain 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters.

This kit has been upgraded to use our Kit 14 chemistry, which includes improved modal raw read sequencing accuracies with high output on our latest nanopore: R10.4.1. The flow cell priming and sequencing reagents are also included as part of the sequencing kit and have been reformulated to be compatible with the Kit 14 chemistry upgrade and R10.4.1 nanopore. Other updates include higher capture rate of gDNA to enable lower flow cell loading amounts, and fuel fix technology, allowing users to run longer experiments without the need for fuel addition during the run.

The kit is supported by the EPI2ME wf-16S workflow, which can be used to analyse data from the 16S Barcoding Kit 24 V14 (SQK-16S114.24) protocol. Deconvolution of barcoded sequencing data is supported by Dorado and EPI2ME which classify the barcode sequence and sort reads into corresponding folders.

Shipping and logistics:

Flow cells and kits are shipped together in an environmentally friendly temperature-controlled shipping box.

Products are shipped to customers within the USA and EU Monday to Thursday. Shipments to Canada, Norway, Korea and Japan are expedited Monday to Wednesday; with Australia and New Zealand leaving our warehouses on a Friday. Shipments to the rest of the world are made on Mondays to allow the full working week for packages to arrive.

The delivery charges are calculated when a quote is raised or during checkout. Once an order is made, the delivery ID and delivery information can be tracked in the Store.

Workflow

The DNA is amplified by PCR using specific 16S primers (27F and 1492R) that contain 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters. The kit contains 24x barcoded primer pairs.

The kit is supported by the EPI2ME wf-16S workflow, that can be used to analyse data from the 16S Barcoding Kit 24 V14 (SQK-16S114.24) protocol. Deconvolution of barcoded sequencing data is supported by Dorado and EPI2ME, which classify the barcode sequence and sort reads into corresponding folders.

16S V14 barcoding workflow

What's in the box

The 16S Barcoding Kit 24 V14 contains 24 unique barcodes and sufficient reagents to generate six sequencing libraries:

SQK-16S114.24 Kit content

NameAcronymCap colourNo. of vialsFill volume per vial (μl)
16S Barcode Primers 01-241-24-2 plates, 3 sets of barcodes per plate15 μl per well
Rapid AdapterRAGreen115
Adapter BufferADBClear1100
AMPure XP BeadsAXPClear cap, light teal label16,000
Elution bufferEBBlack11,500
EDTAEDTABlue1700
Sequencing BufferSBRed1700
Library BeadsLIBPink1600
Library SolutionLISWhite cap, pink label1600
Flow Cell FlushFCFClear cap, light blue label18,000
Flow Cell TetherFCTPurple1200

Note: 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.

3rd party materials

Consumables:

  • LongAmp Hot Start Taq 2X Master Mix (NEB, M0533S)
  • (Optional) Bovine Serum Albumin (BSA) (50 mg/ml) (e.g Invitrogen™ UltraPure™ BSA 50 mg/ml, AM2616)
  • 1.5 ml Eppendorf DNA LoBind tubes
  • 0.2 ml thin-walled PCR tubes
  • Nuclease-free water (e.g. ThermoFisher, AM9937)
  • Freshly prepared 80% ethanol in nuclease-free water
  • Qubit dsDNA HS Assay Kit (Invitrogen, Q32851)
  • Qubit™ Assay Tubes (Invitrogen, Q32856)

Equipment:

  • Ice bucket with ice
  • Microfuge
  • Timer
  • Thermal cycler
  • Magnetic rack
  • Hula mixer (gentle rotator mixer)
  • P1000 pipette with tips
  • P200 pipette and tips
  • P100 pipette and tips
  • P20 pipette and tips
  • P2 pipette and tips
  • Multichannel pipette
  • Qubit fluorometer (or equivalent for QC check)

Optional equipment

  • Agilent Bioanalyzer (or equivalent)
  • Eppendorf 5424 centrifuge (or equivalent)

Compatibility

Kits:

  • Rapid Adapter Auxiliary V14 (EXP-RAA114)
  • Sequencing Auxiliary Vials V14 (EXP-AUX003)
  • Flow Cell Wash Kit (EXP-WSH004)
  • Flow Cell Wash Kit XL (EXP-WSH004-XL)
  • Flow Cell Priming Kit (EXP-FLP004)
  • Flow Cell Priming Kit XL (EXP-FLP004-XL)

Flow cells:

  • FLO-MIN114
  • FLO-FLG114

Devices:

  • MinION Mk1B
  • MinION Mk1C
  • GridION
  • Flongle