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16S Barcoding Kit
SQK-RAB204

Genus-level bacterial identification with barcoding for up to 12 samples.

 Includes a Flow Cell Priming Kit
$650.00

Information

This kit is recommended for users who

  • wish to multiplex samples to reduce price per sample
  • 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.

Barcoding or multiplexing is useful when the amount of data required per sample is less than the total amount of data that can be generated from a single flow cell: it allows a user to pool multiple samples and sequence them together making more efficient use of the flow cell.

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

No barcodes6 barcodes12 barcodes
Flow cell price$500$500$500
Library price$99$99$99
Barcode price-$25$50
Price per sample$599$104$54

The 16S Barcoding Kit offers a method of amplifying and barcoding the ~1500 bp 16S rRNA gene from multiple samples and sequencing them together. 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.

The 16S Barcoding Kit features:

FeatureProperty
Preparation time10 minutes + PCR
Input requirement<10 ng gDNA
PCR RequiredYes
FragmentationNot required
Read lengthFull-length 16S gene (~1.5 kb)
Read type produced1D
Typical throughput*★★☆
Associated protocols16S Barcoding 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 barcodes and 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters. The kit contains 12x barcoded primer pairs.

The kit is supported by the EPI2ME 16S-BLAST workflow, which can be used to analyse data from the 16S protocol. Deconvolution of barcoded sequencing data is supported by Guppy and EPI2ME which classify the barcode sequence and sort reads into corresponding folders.

Shipping and logistics:

Flow cells and kits are shipped together at 2–8°C.

Upon receipt, please place the right product in the right long-term storage location.

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 barcodes and 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters. The kit contains 12 barcoded primer pairs.

The kit is supported by the EPI2ME 16S-BLAST workflow, which can be used to analyse data from the 16S protocol. Deconvolution of barcoded sequencing data is supported by both MinKNOW and Guppy software, which classify the barcode sequence and sort reads into corresponding folders.

RAB v2

Protocols

16S Barcoding Kit

To access the protocol, you will need to register for a Nanopore Community account.

What's in the box

The 16S Barcoding Kit contains 12 unique barcodes and sufficient reagents to generate six sequencing libraries.

2017 12 20 RAB v3 DS

NameAcronymCap colourNo. of vialsFill volume per vial (μl)
Rapid AdapterRAPGreen110
Sequencing TetherSQTPurple110
Loading BeadsLBPink1360
Sequencing BufferSQBRed1300
16S Barcode Primer 1-12FB 16S 01-12Clear1210

The Flow Cell Priming Kit is also supplied. FLP

NameAcronymCap colourNo. of vialsFill volume per vial (μl)
Flush BufferFBBlue61,170
Flush TetherFLTPurple1200

3rd party materials

Consumables

  • Agencourt AMPure XP beads
  • 1.5 ml Eppendorf DNA LoBind tubes
  • 0.2 ml thin-walled PCR tubes
  • Nuclease-free water (e.g. ThermoFisher, cat # AM9937)
  • Freshly prepared 70% ethanol in nuclease-free water
  • LongAmp Taq 2X Master Mix (e.g. NEB M0287)
  • 10 mM Tris-HCl pH 8.0 with 50 mM NaCl

Equipment

  • Microfuge
  • Timer
  • Thermal cycler
  • P1000 pipette and tips
  • P100 pipette and tips
  • P20 pipette and tips
  • P10 pipette and tips
  • P2 pipette and tips

Optional equipment

  • Agilent Bioanalyzer (or equivalent)
  • Qubit fluorometer (or equivalent for QC check)
  • Eppendorf 5424 centrifuge (or equivalent)

Compatibility

The PCR Sequencing Kit can be used together with:

Kits

  • RAP Top-up Kit (EXP-RAP001)
  • Flow Cell Wash Kit (EXP-WSH004)

Flow cells

  • FLO-MINSP6
  • FLO-MIN106D

Devices

  • MinION Mk1B
  • MinION Mk1C
  • GridION