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Ultra-Long DNA Sequencing Kit

SQK-ULK001

$1,200.00

Ultra-Long DNA Sequencing Kit

  • Information

    Ultra-Long DNA Sequencing Kit

    This kit is recommended for users who:

    • are working with ultra-high molecular weight DNA and would like read lengths with an N50 >50 kb

    The Ultra-Long DNA Sequencing Kit offers a means of preparing ultra-high molecular weight (UHMW) DNA for sequencing, which has shown to give N50s >50 kb and reads up to 4+ Mb. The kit is based on transposase chemistry: the transposase simultaneously cleaves template molecules and attaches tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends. The last step is an overnight elution of the DNA library using a Circulomics 5 mm Nanobind disk to remove free adapter and short DNA fragments.

    The kit includes a new type of Rapid Adapter (RAP-F), which is designed such that its consumption of fuel during a sequencing run is reduced significantly (over RAP), resulting in much better maintenance of enzyme speed. As such, the new adapter results in higher sequencing output. 

    Users have achieved the following from an ultra-long preparation for both MinION and PromethION R9.4.1 flow cells is as follows:

    Ultra-Long Example Data 
     
    Feature Property
    Preparation time  ~610 minutes
    Input requirement
    • Tissue culture cells: 6 million
    • Blood: 1–2 ml
    • Gram-positive and -negative bacteria: ~2 ml of OD1 culture
    • Tissue (in development): 5–50 mg (dependent on DNA content)
    PCR Required No
    Fragmentation Transposase-based
    Read length R9.4.1 flow cells: 50–100+ kb N50,
    reads up to 4+ Mb
    Read type produced 1D
    Typical throughput*
    MinION/GridION:R9.4.1 flow cells: 10–20+ Gb
    PromethION:R9.4.1 flow cells: 50–100+ Gb
    Associated protocols Ultra-Long DNA Sequencing Kit (SQK-ULK001)
    Multiplexing options Flow Cell Wash Kit
    Pack size Six reactions where each reaction supports three separate loads of PromethION Flow Cells and six MinION Flow Cells
    Stability Shipped 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.


    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 Ultra-Long DNA Sequencing Kit generates sequencing libraries from extracted ultra-high molecular weight gDNA in ~610 minutes. At the heart of the kit is a transposase which simultaneously cleaves template molecules and attaches tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends, followed by an overnight elution, for example using a Circulomics 5 mm Nanobind disk, to remove free adapter and short DNA fragments.

    2020_11_12_ULK001_v1_DS

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

    The Ultra-Long DNA Sequencing Kit contains sufficient reagents to generate sequencing libraries for six reactions, where each reaction supports three separate loads of PromethION Flow Cells and six MinION Flow Cells.

    SQK-ULK001_kit_contents_v1

    Name Acronym Cap colour No. of tubes Fill volume (μl)
    Rapid Adapter RAP F Green 1 30
    Fragmentation Mix FRA Brown 1 36
    FRA Dilution Buffer FDB Clear 1 1464
    Elution Buffer EB Black 1 1350
    Sequencing Buffer SQB Red 1 1350
    Priming Tether PT Purple 1 540
    Flush Buffer FB White cap, light blue stripe on label 3 7020
    L Fragment Buffer LFB White cap, orange stripe on label 2 6000

     

     

  • 3rd party materials

    Consumables

    • Circulomics Nanobind Big DNA Kit (e.g. CBB, Plant or Tissue)
    • Circulomics Nanobind UL Library Prep Kit (NB-900-601-01)
    • 1.5 ml Eppendorf DNA LoBind tubes

    Equipment

    • Magnetic separator, suitable for 1.5 ml Eppendorf tubes
    • Thermal cycler or heat block at 75°C
    • Vortex mixer
    • Microfuge
    • P1000 pipette and wide-bore pipette tips
    • P200 pipette and wide-bore pipette tips
    • P200 pipette and tips
    • Ice bucket with ice
    • Timer

    Optional Equipment

    • Qubit fluorometer (or equivalent for QC check)
  • Compatibility

    The Ultra-Long DNA Sequencing Kit can be used together with:

    Kits

    • Flow Cell Wash Kit (EXP-WSH004)

    Flow cells

    • FLO-MINSP6
    • FLO-MIN106D
    • FLO-MIN111
    • FLO-PRO002

    Devices

    • MinION Mk1B
    • MinION Mk1C
    • GridION Mk1
    • PromethION P24/48
Information

Ultra-Long DNA Sequencing Kit

This kit is recommended for users who:

  • are working with ultra-high molecular weight DNA and would like read lengths with an N50 >50 kb

The Ultra-Long DNA Sequencing Kit offers a means of preparing ultra-high molecular weight (UHMW) DNA for sequencing, which has shown to give N50s >50 kb and reads up to 4+ Mb. The kit is based on transposase chemistry: the transposase simultaneously cleaves template molecules and attaches tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends. The last step is an overnight elution of the DNA library using a Circulomics 5 mm Nanobind disk to remove free adapter and short DNA fragments.

The kit includes a new type of Rapid Adapter (RAP-F), which is designed such that its consumption of fuel during a sequencing run is reduced significantly (over RAP), resulting in much better maintenance of enzyme speed. As such, the new adapter results in higher sequencing output. 

Users have achieved the following from an ultra-long preparation for both MinION and PromethION R9.4.1 flow cells is as follows:

Ultra-Long Example Data 
 
Feature Property
Preparation time  ~610 minutes
Input requirement
  • Tissue culture cells: 6 million
  • Blood: 1–2 ml
  • Gram-positive and -negative bacteria: ~2 ml of OD1 culture
  • Tissue (in development): 5–50 mg (dependent on DNA content)
PCR Required No
Fragmentation Transposase-based
Read length R9.4.1 flow cells: 50–100+ kb N50,
reads up to 4+ Mb
Read type produced 1D
Typical throughput*
MinION/GridION:R9.4.1 flow cells: 10–20+ Gb
PromethION:R9.4.1 flow cells: 50–100+ Gb
Associated protocols Ultra-Long DNA Sequencing Kit (SQK-ULK001)
Multiplexing options Flow Cell Wash Kit
Pack size Six reactions where each reaction supports three separate loads of PromethION Flow Cells and six MinION Flow Cells
Stability Shipped 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.


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 Ultra-Long DNA Sequencing Kit generates sequencing libraries from extracted ultra-high molecular weight gDNA in ~610 minutes. At the heart of the kit is a transposase which simultaneously cleaves template molecules and attaches tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends, followed by an overnight elution, for example using a Circulomics 5 mm Nanobind disk, to remove free adapter and short DNA fragments.

2020_11_12_ULK001_v1_DS

Protocols
What's in the box

The Ultra-Long DNA Sequencing Kit contains sufficient reagents to generate sequencing libraries for six reactions, where each reaction supports three separate loads of PromethION Flow Cells and six MinION Flow Cells.

SQK-ULK001_kit_contents_v1

Name Acronym Cap colour No. of tubes Fill volume (μl)
Rapid Adapter RAP F Green 1 30
Fragmentation Mix FRA Brown 1 36
FRA Dilution Buffer FDB Clear 1 1464
Elution Buffer EB Black 1 1350
Sequencing Buffer SQB Red 1 1350
Priming Tether PT Purple 1 540
Flush Buffer FB White cap, light blue stripe on label 3 7020
L Fragment Buffer LFB White cap, orange stripe on label 2 6000

 

 

3rd party materials

Consumables

  • Circulomics Nanobind Big DNA Kit (e.g. CBB, Plant or Tissue)
  • Circulomics Nanobind UL Library Prep Kit (NB-900-601-01)
  • 1.5 ml Eppendorf DNA LoBind tubes

Equipment

  • Magnetic separator, suitable for 1.5 ml Eppendorf tubes
  • Thermal cycler or heat block at 75°C
  • Vortex mixer
  • Microfuge
  • P1000 pipette and wide-bore pipette tips
  • P200 pipette and wide-bore pipette tips
  • P200 pipette and tips
  • Ice bucket with ice
  • Timer

Optional Equipment

  • Qubit fluorometer (or equivalent for QC check)
Compatibility

The Ultra-Long DNA Sequencing Kit can be used together with:

Kits

  • Flow Cell Wash Kit (EXP-WSH004)

Flow cells

  • FLO-MINSP6
  • FLO-MIN106D
  • FLO-MIN111
  • FLO-PRO002

Devices

  • MinION Mk1B
  • MinION Mk1C
  • GridION Mk1
  • PromethION P24/48
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