This kit is recommended for users who:

• require a short preparation time
• have limited access to laboratory equipment.

The Rapid Sequencing Kit generates 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 tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends.

The Rapid Sequencing Kit features:

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 Sequencing Kit.

Typical throughput achieved with the Rapid Sequencing Kit compared with the Ligation Sequencing Kit.

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

Further considerations:

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

Compare Sequencing Kits

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

The Rapid Sequencing Kit generates 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 tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends.

Example protocol using Rapid Sequencing Kit (SQK-RAD004) - for illustration purposes only

Safety Data Sheet for Rapid Sequencing Kit

Safety Data Sheet for Flow Cell Priming Kit

The Rapid Sequencing Kit contains sufficient reagents to generate six sequencing libraries.

The Flow Cell Priming Kit Expansion Pack is also supplied.

PCR-free multiplexing with barcoding for up to 12 samples is available with an alternative purchase: Rapid Barcoding Kit (SQK-RBK004).

Consumables

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

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

• Qubit fluorometer (or equivalent for QC check)

Additional materials for:

Rapid whole genome amplification

• Qiagen REPLI-g UltraFast Mini Kit
• Phosphate buffered saline (PBS)

The Rapid 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
• GridION X5
• PromethION