Sequence RNA molecules directly and preserve base modifications.

This uses Kit 9 chemistry.

Includes a Flow Cell Priming Kit

This product is scheduled to be discontinued in March 2024 and removed from the store January 2024. The recommended replacement is SQK-RNA004

£545.00

5Legacy

These Products are nearing their end of life and will soon be discontinued
Users need to contact customer support to discuss project completion timelines
Warranty periods of 3 months or more
Product discontinuation within 3-6 months
Lead time visible in store
Product subject to availability

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

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.

The Direct RNA Sequencing Kit features:

Feature	Property
Preparation time	105 minutes
Input requirement	500 ng total RNA or 50 ng poly(A)+ RNA
Reverse transcription required	Optional
PCR required	No
Read length	Equal to RNA length
Kit chemistry	Kit 9
Typical number of reads	1 million full-length per flow cell on MinION/GridION; 8 million per flow cell on PromethION
Associated protocols	• Direct RNA sequencing - control experiment (SQK-RNA002) • Direct RNA sequencing (SQK-RNA002) • Direct RNA sequencing - sequence-specific (SQK-RNA002)
Multiplexing options	Flow Cell Wash Kit
Pack size	6 reactions
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.

Further considerations:

The direct RNA sequencing chemistry is new and both accuracy and throughput will continue to improve with future 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 output per run.

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 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 output.

2019 02 18 RNA002 v1 DS

Protocols

• Direct RNA sequencing - control experiment (SQK-RNA002)
• Direct RNA sequencing (SQK-RNA002)
• Direct RNA sequencing - sequence-specific (SQK-RNA002)

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

Safety and legal

Safety Data Sheet for Direct RNA Sequencing Kit (SQK-RNA002)

Safety Data Sheet for Flow Cell Priming Kit

What's in the box

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

dRNA tubes

Name	Acronym	Cap colour	No. of vials	Fill volume per vial (μl)
RT Adapter	RTA	Blue	1	10
RNA Adapter Mix	RMX	Green	1	45
RNA Calibrant Strand	RCS	Yellow	1	25
Wash Buffer	WSB	Orange	2	1,200
Elution Buffer	ELB	Black	1	300
RNA Running Buffer	RRB	Red	1	600

The Flow Cell Priming Kit is also supplied.

FLP

Name	Acronym	Cap colour	No. of vials	Fill volume per vial (μl)
Flush Buffer	FB	Blue	6	1,170
Flush Tether	FLT	Purple	1	200

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-WSH004)

Flow cells

FLO-MIN106D
FLO-PRO002

Devices

MinION Mk1B
MinION Mk1C
GridION
PromethION

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