Information
This kit is recommended for users who
- Wish to multiplex samples to reduce price per sample
- Want to do 16S sequencing
- Are interested in genus level bacterial identification
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.
This is a Legacy product
This kit is soon to be discontinued and we recommend all customers to upgrade to the latest chemistry for their relevant kit which is available on the Store. If customers require further support for any ongoing critical experiments using a Legacy product, please contact Customer Support via email: support@nanoporetech.com.
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. Please note, this is an example and pricing depends on the kit purchased.
| No barcodes | 6 barcodes | 12 barcodes |
|---|
| Flow cell price | $500 | $500 | $500 |
| Library price | $99 | $99 | $99 |
| Barcode price | - | $25 | $50 |
| Price per sample | $599 | $104 | $54 |
The 16S Barcoding Kit offers a method of amplifying and barcoding the ~1500 bp 16S rRNA gene from multiple samples and sequencing them together. By narrowing down to a specific region of interest, a user can see all the organisms in the sample without sequencing unnecessary regions of the genome, making the identification quicker and more economical.
The 16S Barcoding Kit features:
| Feature | Property |
|---|
| Preparation time | 10 minutes + PCR |
| Input requirement | <10 ng gDNA per sample |
| PCR Required | Yes |
| Fragmentation | Not required |
| Read length | Full-length 16S gene (~1.5 kb) |
| Kit chemistry | Kit 9 |
| Associated protocols | 16S Barcoding 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. |
The DNA is amplified by PCR using specific 16S primers (27F and 1492R) that contain barcodes and 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters. The kit contains 12x barcoded primer pairs.
The kit is supported by the EPI2ME 16S-BLAST workflow, which can be used to analyse data from the 16S protocol. Deconvolution of barcoded sequencing data is supported by Guppy and EPI2ME which classify the barcode sequence and sort reads into corresponding folders.
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 DNA is amplified by PCR using specific 16S primers (27F and 1492R) that contain barcodes and 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters. The kit contains 12 barcoded primer pairs.
The kit is supported by the EPI2ME 16S-BLAST workflow, which can be used to analyse data from the 16S protocol. Deconvolution of barcoded sequencing data is supported by both MinKNOW and Guppy software, which classify the barcode sequence and sort reads into corresponding folders.
Protocols
Rapid sequencing amplicons - 16S barcoding (SQK-RAB204)
To access the protocol, you will need to register for a Nanopore Community account.
What's in the box
The 16S Barcoding Kit contains 12 unique barcodes and sufficient reagents to generate six sequencing libraries.
| Name | Acronym | Cap colour | No. of vials | Fill volume per vial (μl) |
|---|
| Rapid Adapter | RAP | Green | 1 | 10 |
| Sequencing Tether | SQT | Purple | 1 | 10 |
| Loading Beads | LB | Pink | 1 | 360 |
| Sequencing Buffer | SQB | Red | 1 | 300 |
| 16S Barcode Primer 1-12 | FB 16S 01-12 | Clear | 12 | 10 |
The Flow Cell Priming Kit is also supplied.
| 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 |
Compatibility
The PCR Sequencing Kit can be used together with:
Kits
- RAP Top-up Kit (EXP-RAP001)
- Flow Cell Wash Kit (EXP-WSH004)
Flow cells
Devices
- MinION Mk1B
- MinION Mk1C
- GridION