Express delivery and free returns within 30 days

Rapid Barcoding Kit 96

Simple and rapid library preparation, with barcoding for up to 96 gDNA samples. We recommend using our most recent version: Rapid Barcoding Kit 96 V14 (SQK-RBK114.96).

This uses Kit 10 chemistry.

SKU SQK-RBK110.96 Categorías ,
Share it :

Descripción

Information

This kit is recommended for users who:

  • Wish to multiplex samples to reduce price per sample
  • Need a PCR-free method of multiplexing to preserve additional information such as base modifications
  • Require a short preparation time
  • Have limited access to laboratory equipment

We recommend using our most recent version: Rapid Barcoding Kit 96 V14 (SQK-RBK114.96).

Please note that to use this kit, you will need to purchase additional 3rd party materials: see the «3rd Party Materials» tab for more detail.

Barcoding or multiplexing is useful when the amount of data required per sample is less than the total amount of data that can be generated from a single flow cell: it allows a user to pool multiple samples and sequence them together, making more efficient use of the flow cell.

Multiplexing samples onto one flow cell can reduce the cost per sample for a user. Below are some worked examples:

No barcodes 6 barcodes 12 barcodes 96 barcodes
Flow cell price $500 $500 $500 $500
Library price $99 $99 $99 $99
Price per sample $599 $104 $54 $5

The Rapid Barcoding Kit 96 features:

Feature Property
Preparation time 60 minutes
Input requirement 50 ng gDNA (>30 kb) per sample
PCR Required No
Fragmentation Transposase-based
Read length Random distribution, dependent on input fragment length
Kit chemistry Kit 10
Associated protocols • Rapid sequencing gDNA – barcoding (SQK-RBK110.96)

• PCR tiling of SARS-CoV-2 virus – rapid barcoding (SQK-RBK110.96)

Multiplexing options • Rapid Barcoding Kit 96

• Flow Cell Wash Kit

Pack size Up to 12 reactions:

Options: 12 libraries containing 24 barcodes, 6 libraries containing 48 barcodes, 3 libraries containing 96 barcode

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.

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, some very long reads can be achieved with this kit, despite the required transposase fragmentation. However, 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 and therefore removes PCR bias and retains base modification information which can be analysed using tools developed in the Nanopore Community.

Deconvolution of barcoded sequencing data is supported by MinKNOW and Guppy software, which classify the barcode sequence and sort reads into corresponding folders.

Further considerations:

The Rapid Barcoding Kit recommends an input of 50 ng gDNA per sample, and is optimised for samples which contain long fragments (>30 kb). Addition of less than 50 ng, or shorter fragments could compromise sequencing throughput and read length. For samples where only lower inputs are possible, the Rapid PCR Barcoding Kit is available.

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 Barcoding Kit 96 generates barcoded sequencing libraries from extracted gDNA in 60 minutes using a simple protocol. At the heart of the kit is a transposase which simultaneously cleaves template molecules and attaches barcoded tags to the cleaved ends: there are 96 unique barcoded tags in the kit. Barcoded samples are pooled, purified using SPRI beads, and Rapid Sequencing Adapters are then added to the tagged ends.

SQK-RBK110.96 gDNA workflow v1

Protocols

• Rapid sequencing gDNA – barcoding (SQK-RBK110.96)

• PCR tiling of SARS-CoV-2 virus – rapid barcoding (SQK-RBK110.96)

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

What’s in the box

Q-SQK-RBK110.96

Name Acronym Cap colour No. of vials Fill volume per vial (µl)
Rapid Barcode plate RB96 3 plates 8 µl per well
AMPure XP Beads AXP Brown 3 1,200
Sequencing Buffer II SBII Red 1 500
Rapid Adapter F RAP-F Green 1 25
Elution Buffer EB Black 1 500
Loading Beads II LBII Pink 1 360
Loading Solution LS White cap, pink label 1 400
Flush Tether FLT Purple 1 400
Flush Buffer FB White 1 bottle 15,500

This product contains AMPure XP reagent manufactured by Beckman Coulter, Inc.

3rd party materials

Consumables

  • 1.5 ml Eppendorf DNA LoBind tubes
  • 2 ml Eppendorf DNA LoBind tubes
  • 0.2 ml thin-walled PCR tubes
  • Eppendorf twin.tec® PCR plate 96 LoBind, semi-skirted (Cat # 0030129504) with heat seals
  • Nuclease-free water (e.g. ThermoFisher, cat # AM9937)
  • Freshly-prepared 80% ethanol in nuclease-free water

Equipment

  • Ice bucket with ice
  • Microplate centrifuge, e.g. Fisherbrand™ Mini Plate Spinner Centrifuge (Fisher Scientific, #11766427)
  • Timer
  • Thermal cycler or heat block at 30°C and 80°C
  • Magnetic rack
  • Hula mixer (gentle rotator mixer)
  • P1000 pipette and tips
  • P200 pipette and tips
  • P100 pipette and tips
  • P20 pipette and tips
  • P2 pipette and tips
  • Multichannel pipette

Optional equipment

  • Qubit fluorometer (or equivalent for QC check)

Compatibility

The Rapid Barcoding Kit 96 can be used together with:

Kits

  • Flow Cell Wash Kit (EXP-WSH004)
  • Sequencing Auxiliary Vials (EXP-AUX002)

Flow cells

  • FLO-MIN106D
  • FLO-PRO002
  • FLO-FLG001

Devices

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

Valoraciones

No hay valoraciones aún.

Sé el primero en valorar “Rapid Barcoding Kit 96”

Tu dirección de correo electrónico no será publicada. Los campos obligatorios están marcados con *