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Rapid Sequencing Kit V14

A simple and rapid library preparation method for genomic DNA optimised for speed with improved modal raw read accuracy.

This is an Early Access product

This uses our latest Kit 14 chemistry

SKU SQK-RAD114 Categorías ,
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Descripción

Information

This kit is recommended for users who:

  • Require a short preparation time
  • Have limited access to laboratory equipment

This is an Early Access product.

For more information about our Early Access programmes, please see this article on product release phases.

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 Rapid Sequencing Kit V14 features:

Features Property
Preparation time 10 minutes
Input requirement 100 ng of gDNA
PCR requirement No
Read length Random distribution, dependent on input fragment length
Kit chemistry Kit 14 (V14)
Associated protocols Rapid Sequencing Kit V14 – gDNA (SQK-RAD114)

Rapid Sequencing Kit V14 – Lambda control (SQK-RAD114)

Pack size 6 reactions
Storage Shipped at 2–8°C

Long-term storage -20°C

The warranty for this Early Access product is 3 months from receipt by the customer

The Rapid Sequencing Kit V14 (SQK-RAD114) is a standalone kit optimised for speed and simplicity to sequence libraries using a PCR-free method. The kit uses a transposase which simultaneously cleaves template molecules and attaches tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends for sequencing on a flow cell.

This kit has been updated to use our newest Kit 14 chemistry which includes improved modal raw read sequencing accuracies with higher output on our latest nanopore: R10.4.1. The flow cell priming and sequencing reagents are now included as part of the sequencing kit and have been reformulated to be compatible with the chemistry upgrade and R10.4.1 nanopore.

Kit 14 also includes previous updates such as the higher capture rate of DNA to enable lower flow cell loading amounts, and fuel fix technology, allowing users to run longer experiments without the need for fuel addition during the run.

Note: This kit is able to generate Q20+ and duplex data but has been optimised for speed and simplicity. If users require high accuracy, we recommend using our Ligation Sequencing Kit V14 (SQK-LSK114).

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-up steps) 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.

The workflow is PCR-free, removing the 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 100 ng gDNA, and is optimised for samples which contain long fragments (>30 kb). Addition of less than 100 ng, or the presence of shorter fragments could compromise sequencing output and read length.

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 Rapid Sequencing Kit V14 generates sequencing libraries from extracted gDNA in 10 minutes using a simple two-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.

Protocols

• Rapid Sequencing Kit V14 – gDNA (SQK-LSK114)

• Rapid Sequencing Kit V14 – Lambda control (SQK-LSK114)

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

What’s in the box

The Rapid Sequencing Kit V14 (SQK-RAD114) contains sufficient reagents for six reactions.

Note: We are in the process of reformatting our kits with single-use tubes into a bottle format.

Single-use tubes format:

RAD114 tubes

Name Acronym Cap colour No. of vials Fill volume per vial (µl)
Fragmentation Mix FRA Orange 1 10
Rapid Adapter RA Green 1 15
Adapter Buffer ADB Clear 1 100
Library Beads LIB Pink 1 600
Library Solution LIS White cap, pink label 1 600
Sequencing Buffer SB Red 1 700
Flow Cell Flush FCF Blue 6 1170
Flow Cell Tether FCT Purple 1 200

Bottle format:

RAD114 bottle

Name Acronym Cap colour No. of vials Fill volume per vial (µl)
Fragmentation Mix FRA Orange 1 10
Rapid Adapter RA Green 1 15
Adapter Buffer ADB Clear 1 100
Library Beads LIB Pink 1 600
Library Solution LIS White cap, pink label 1 600
Sequencing Buffer SB Red 1 700
Flow Cell Flush FCF Blue 1 8,000
Flow Cell Tether FCT 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)
  • (Optional) Bovine Serum Albumin (BSA) (50 mg/ml) (e.g Invitrogen™ UltraPure™ BSA 50 mg/ml, cat# AM2616)

Equipment

  • Microfuge
  • P1000 pipette and tips
  • P100 pipette and tips
  • P20 pipette and tips
  • P10 pipette and tips
  • P2 pipette and tips
  • Timer
  • Thermal cycler or heat block at 30°C and 80°C

Optional equipment

  • Qubit fluorometer (or equivalent for QC check)

Compatibility

The Rapid Sequencing Kit V14 can be used together with:

Kits:

  • Flow Cell Wash Kit (EXP-WSH004)
  • Control Expansion (EXP-CTL001)
  • Sequencing Auxiliary Expansion V14 (EXP-AUX003)
  • Flow Cell Priming Kit (EXP-FLP004)
  • Flow Cell Priming Kit XL (EXP-FLP004-XL)
  • Rapid Adapter Auxiliary V14 (EXP-RAA114)

Flow cells:

R10.4.1:

  • FLO-MIN114
  • FLO-PRO114M
  • FLO-FLG114

Devices:

  • Flongle
  • MinION Mk1B
  • MinION Mk1C
  • GridION
  • PromethION

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