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Direct RNA Sequencing Kit

SQK-RNA002

$599.00

Sequence RNA molecules directly and preserve base modifications

Includes a Flow Cell Priming Kit

Poly-A+ RNA 500 ng 105 mins Direct RNA RNA base mods
  • Information

    This kit is highly recommended for users who:

    • would like to remove RT or PCR bias
    • have transcripts that are difficult to reverse transcribe
    • are exploring attributes of native RNA such as modified bases

     

    The Direct RNA Sequencing Kit features:

    Feature Property
    Preparation time 105 minutes
    Input requirement 500 ng poly-A+ RNA
    Reverse transcription required Optional
    PCR required No
    Read length Equal to RNA length
    Read type produced 1D
    Typical throughput* ••
    Typical number of reads 1 million full-length per flow cell on MinION/GridION; 3 million per flow cell on PromethION
    Associated protocols

    Direct RNA Sequencing 

    Sequence-specific direct RNA sequencing

    Multiplexing options

    Barcoding kits in development

    Flow Cell Wash Kit

    Pack size 6 reactions
    Stability

    Shipped at  2-8° C

    Long-term storage  -20° C

     

    Description

    The Direct RNA Sequencing kit (SQK-RNA002) is used to prepare any RNA with a 3’ polyA tail for 1D sequencing on the Oxford Nanopore sequencing devices. Possible input RNA includes eukaryotic mRNA, viral RNA with a polyA tail, or any RNA prepared with a polyA-tailing kit. For RNA without a polyA tail, users can follow simple instructions to design their own custom adapter to ligate a specific terminal 3’ sequence like the 3’ of tRNA or 16S rRNA. The kit requires 500 ng of input RNA.

     

    In contrast to cDNA sequencing protocols, which are used to prepare cDNA copies of RNA for sequencing, the Direct RNA kit is used to prepare RNA itself for sequencing. In Direct RNA Sequencing, only the native RNA passes through the nanopore sensor thus the read length reflects the length of the RNA molecules in the sample.

    The Direct RNA Sequencing protocol contains an optional (but recommended) reverse transcription step and though the cDNA strand generated in this step is not sequenced, it significantly improves sequencing throughput. 

    Further Considerations

    The Direct RNA Sequencing kit is new and both accuracy and throughput will continue to improve with coming kit versions. In applications where Direct RNA sequencing’s key advantages (detection of modified bases and reduced bias) are not important, users may consider one of Oxford Nanopore’s cDNA kits because they currently provide the highest throughput per run.

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

  • Protocols

    Example protocol using Direct RNA Sequencing Kit - for illustration purposes only

  • Safety and legal
  • What's in the box

    The Direct RNA Sequencing Kit contains sufficient reagents to generate six sequencing libraries

     

    The Flow Cell Priming 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)
    • Freshly prepared 70% ethanol in nuclease-free water
    • SuperScript III Reverse Transcriptase (Thermo Fisher Scientific, cat # 18080044)
    • 10 mM dNTP solution (e.g. NEB cat # N0447)
    • NEBNext® Quick Ligation Reaction Buffer (NEB B6058)
    • T4 DNA Ligase 2M U/ml (NEB M0202)
    • Agencourt RNAClean XP beads
    • Qubit RNA HS Assay Kit (ThermoFisher cat # Q32852)
    • Qubit dsDNA HS Assay Kit (ThermoFisher cat #  Q32851)

    Equipment

    • Hula mixer (gentle rotator mixer)
    • Magnetic rack
    • Microfuge
    • Vortex mixer
    • Ice bucket with ice
    • 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 Direct RNA Sequencing Kit can be used together with:

    Kits

    • Flow Cell Wash Kit (EXP-WSH003)

    Flow cells

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

    Devices

    • MinION Mk 1B
    • MinION Mk 1C
    • GridION Mk1
    • PromethION alpha-beta/P24/P48
Information

This kit is highly recommended for users who:

  • would like to remove RT or PCR bias
  • have transcripts that are difficult to reverse transcribe
  • are exploring attributes of native RNA such as modified bases

 

The Direct RNA Sequencing Kit features:

Feature Property
Preparation time 105 minutes
Input requirement 500 ng poly-A+ RNA
Reverse transcription required Optional
PCR required No
Read length Equal to RNA length
Read type produced 1D
Typical throughput* ••
Typical number of reads 1 million full-length per flow cell on MinION/GridION; 3 million per flow cell on PromethION
Associated protocols

Direct RNA Sequencing 

Sequence-specific direct RNA sequencing

Multiplexing options

Barcoding kits in development

Flow Cell Wash Kit

Pack size 6 reactions
Stability

Shipped at  2-8° C

Long-term storage  -20° C

 

Description

The Direct RNA Sequencing kit (SQK-RNA002) is used to prepare any RNA with a 3’ polyA tail for 1D sequencing on the Oxford Nanopore sequencing devices. Possible input RNA includes eukaryotic mRNA, viral RNA with a polyA tail, or any RNA prepared with a polyA-tailing kit. For RNA without a polyA tail, users can follow simple instructions to design their own custom adapter to ligate a specific terminal 3’ sequence like the 3’ of tRNA or 16S rRNA. The kit requires 500 ng of input RNA.

 

In contrast to cDNA sequencing protocols, which are used to prepare cDNA copies of RNA for sequencing, the Direct RNA kit is used to prepare RNA itself for sequencing. In Direct RNA Sequencing, only the native RNA passes through the nanopore sensor thus the read length reflects the length of the RNA molecules in the sample.

The Direct RNA Sequencing protocol contains an optional (but recommended) reverse transcription step and though the cDNA strand generated in this step is not sequenced, it significantly improves sequencing throughput. 

Further Considerations

The Direct RNA Sequencing kit is new and both accuracy and throughput will continue to improve with coming kit versions. In applications where Direct RNA sequencing’s key advantages (detection of modified bases and reduced bias) are not important, users may consider one of Oxford Nanopore’s cDNA kits because they currently provide the highest throughput per run.

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

Protocols

Example protocol using Direct RNA Sequencing Kit - for illustration purposes only

What's in the box

The Direct RNA Sequencing Kit contains sufficient reagents to generate six sequencing libraries

 

The Flow Cell Priming 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)
  • Freshly prepared 70% ethanol in nuclease-free water
  • SuperScript III Reverse Transcriptase (Thermo Fisher Scientific, cat # 18080044)
  • 10 mM dNTP solution (e.g. NEB cat # N0447)
  • NEBNext® Quick Ligation Reaction Buffer (NEB B6058)
  • T4 DNA Ligase 2M U/ml (NEB M0202)
  • Agencourt RNAClean XP beads
  • Qubit RNA HS Assay Kit (ThermoFisher cat # Q32852)
  • Qubit dsDNA HS Assay Kit (ThermoFisher cat #  Q32851)

Equipment

  • Hula mixer (gentle rotator mixer)
  • Magnetic rack
  • Microfuge
  • Vortex mixer
  • Ice bucket with ice
  • 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 Direct RNA Sequencing Kit can be used together with:

Kits

  • Flow Cell Wash Kit (EXP-WSH003)

Flow cells

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

Devices

  • MinION Mk 1B
  • MinION Mk 1C
  • GridION Mk1
  • PromethION alpha-beta/P24/P48
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