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Rapid Barcoding Kit

SQK-RBK004

$650.00

Simple and rapid library preparation, with barcoding for up to 12 gDNA samples

Includes a Flow Cell Priming Kit

gDNA 400 ng 10 mins No PCR Speed / simplicity
  • Information

    This kit is recommended for users who:

    • wish to multiplex samples to reduce price per sample
    • need a PCR-free method of multiplexing to preserve additional information such as base modifications
    • require a short preparation time
    • have limited access to laboratory equipment.

    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 Rapid Barcoding Kit generates barcoded sequencing libraries from extracted gDNA in 10 minutes using a simple 2-step protocol. At the heart of the kit is a transposase which simultaneously cleaves template molecules and attaches barcoded tags to the cleaved ends: there are 12 unique barcoded tags in the kit. Barcoded samples are pooled and Rapid Sequencing Adapters are then added to the tagged ends.

    The Rapid Barcoding Kit features:

    Feature Property
    Preparation time 10 minutes
    Input requirement 400 ng of high molecular weight gDNA (>30 kb)
    PCR Required No
    Fragmentation Transposase-based
    Read length Random distribution, dependent on input fragment length
    Read type produced 1D
    Typical throughput* ••
    Associated protocols

    Rapid Barcoding Kit

    Flow Cell Wash Kit

    Pack size 6 reactions
    Stability

    Shipped at  2–8° C

    Long-term storage  -20° C

     

    The kit is optimised for simplicity and speed, rather than for obtaining maximum throughput. Due to the simple nature of the workflow and the fact that little sample manipulation is required (e.g. minimal pipetting steps and no clean-ups), some very long reads can be achieved with this kit, despite the required transposase fragmentation. However, in order for long reads to be observed in sequencing, long fragments need to be present in the sample in the first place.

     

    Typical read length histogram when preparing an E.coli library with the Rapid Barcoding Kit.

    The workflow is PCR-free and therefore removes PCR bias and retains base modification information which can be analysed using tools developed in the Nanopore Community.

    Deconvolution of barcoded sequencing data is supported by Albacore and EPI2ME which classify the barcode sequence and sort reads into corresponding folders.

    Further considerations:

    The Rapid Barcoding Kit recommends a total input of 400 ng gDNA, and is optimised for samples which contain long fragments (>30 kb). Addition of less than 400 ng, or shorter fragments could compromise sequencing throughput and read length. For samples where only lower inputs are possible, the Low Input by PCR Barcoding Kit and Rapid Low Input by PCR Barcoding Kit are available.

    Additional Rapid Sequencing Adapter (RAP) can be purchased (EXP-RAP001) to support the use of the Rapid Barcoding Kit.

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

  • Workflow

    The Rapid Barcoding Kit generates barcoded sequencing libraries from extracted gDNA in 10 minutes using a simple 2-step protocol. At the heart of the kit is a transposase which simultaneously cleaves template molecules and attaches barcoded tags to the cleaved ends: there are 12 unique barcoded tags in the kit. Barcoded samples are pooled and Rapid Sequencing Adapters are then added to the tagged ends.

  • Protocols
  • Safety and legal
  • What's in the box

    The Rapid Barcoding Kit contains twelve unique barcodes and sufficient reagents to generate six sequencing libraries.

     

    The Flow Cell Priming Kit Expansion Pack is also supplied.

    EXP-FLP002

  • 3rd party materials

    Consumables

    • 1.5 ml Eppendorf DNA LoBind tubes
    • 0.2 ml thin-walled PCR tubes
    • Nuclease-free water (e.g. ThermoFisher, cat # AM9937)

    Optional consumables

    • Agencourt AMPure XP beads
    • Freshly-prepared 70% ethanol in nuclease-free water
    • 10 mM Tris-HCl pH 8.0 with 50 mM NaCl

    Equipment

    • Microfuge
    • Vortex mixer
    • Thermal cycler at 30° C and 80° C
    • P1000 pipette and tips
    • P100 pipette and tips
    • P20 pipette and tips
    • P10 pipette and tips
    • P2 pipette and tips
    • Timer

    Optional Equipment

    • Agilent Bioanalyzer (or equivalent)
    • Qubit fluorometer (or equivalent for QC check)
    • Eppendorf 5424 centrifuge (or equivalent)
    • Magnetic rack
    • Hula mixer (gentle rotator mixer)

     

  • Compatibility

    The Rapid Barcoding Kit can be used together with:

    Kits

    • RAP Top-up Kit (EXP-RAP001)
    • Flow Cell Wash Kit (EXP-WSH003)

    Flow cells

    • FLO-MINSP6
    • FLO-MIN106D

    Devices

    • MinION Mk 1B
    • MinION Mk 1C
    • GridION Mk1
Information

This kit is recommended for users who:

  • wish to multiplex samples to reduce price per sample
  • need a PCR-free method of multiplexing to preserve additional information such as base modifications
  • require a short preparation time
  • have limited access to laboratory equipment.

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 Rapid Barcoding Kit generates barcoded sequencing libraries from extracted gDNA in 10 minutes using a simple 2-step protocol. At the heart of the kit is a transposase which simultaneously cleaves template molecules and attaches barcoded tags to the cleaved ends: there are 12 unique barcoded tags in the kit. Barcoded samples are pooled and Rapid Sequencing Adapters are then added to the tagged ends.

The Rapid Barcoding Kit features:

Feature Property
Preparation time 10 minutes
Input requirement 400 ng of high molecular weight gDNA (>30 kb)
PCR Required No
Fragmentation Transposase-based
Read length Random distribution, dependent on input fragment length
Read type produced 1D
Typical throughput* ••
Associated protocols

Rapid Barcoding Kit

Flow Cell Wash Kit

Pack size 6 reactions
Stability

Shipped at  2–8° C

Long-term storage  -20° C

 

The kit is optimised for simplicity and speed, rather than for obtaining maximum throughput. Due to the simple nature of the workflow and the fact that little sample manipulation is required (e.g. minimal pipetting steps and no clean-ups), some very long reads can be achieved with this kit, despite the required transposase fragmentation. However, in order for long reads to be observed in sequencing, long fragments need to be present in the sample in the first place.

 

Typical read length histogram when preparing an E.coli library with the Rapid Barcoding Kit.

The workflow is PCR-free and therefore removes PCR bias and retains base modification information which can be analysed using tools developed in the Nanopore Community.

Deconvolution of barcoded sequencing data is supported by Albacore and EPI2ME which classify the barcode sequence and sort reads into corresponding folders.

Further considerations:

The Rapid Barcoding Kit recommends a total input of 400 ng gDNA, and is optimised for samples which contain long fragments (>30 kb). Addition of less than 400 ng, or shorter fragments could compromise sequencing throughput and read length. For samples where only lower inputs are possible, the Low Input by PCR Barcoding Kit and Rapid Low Input by PCR Barcoding Kit are available.

Additional Rapid Sequencing Adapter (RAP) can be purchased (EXP-RAP001) to support the use of the Rapid Barcoding Kit.

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

Workflow

The Rapid Barcoding Kit generates barcoded sequencing libraries from extracted gDNA in 10 minutes using a simple 2-step protocol. At the heart of the kit is a transposase which simultaneously cleaves template molecules and attaches barcoded tags to the cleaved ends: there are 12 unique barcoded tags in the kit. Barcoded samples are pooled and Rapid Sequencing Adapters are then added to the tagged ends.

What's in the box

The Rapid Barcoding Kit contains twelve unique barcodes and sufficient reagents to generate six sequencing libraries.

 

The Flow Cell Priming Kit Expansion Pack is also supplied.

EXP-FLP002

3rd party materials

Consumables

  • 1.5 ml Eppendorf DNA LoBind tubes
  • 0.2 ml thin-walled PCR tubes
  • Nuclease-free water (e.g. ThermoFisher, cat # AM9937)

Optional consumables

  • Agencourt AMPure XP beads
  • Freshly-prepared 70% ethanol in nuclease-free water
  • 10 mM Tris-HCl pH 8.0 with 50 mM NaCl

Equipment

  • Microfuge
  • Vortex mixer
  • Thermal cycler at 30° C and 80° C
  • P1000 pipette and tips
  • P100 pipette and tips
  • P20 pipette and tips
  • P10 pipette and tips
  • P2 pipette and tips
  • Timer

Optional Equipment

  • Agilent Bioanalyzer (or equivalent)
  • Qubit fluorometer (or equivalent for QC check)
  • Eppendorf 5424 centrifuge (or equivalent)
  • Magnetic rack
  • Hula mixer (gentle rotator mixer)

 

Compatibility

The Rapid Barcoding Kit can be used together with:

Kits

  • RAP Top-up Kit (EXP-RAP001)
  • Flow Cell Wash Kit (EXP-WSH003)

Flow cells

  • FLO-MINSP6
  • FLO-MIN106D

Devices

  • MinION Mk 1B
  • MinION Mk 1C
  • GridION Mk1

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