Information
This kit is recommended for users who:
- 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.
Multiplexing samples onto one flow cell can reduce the cost per sample for a user. Below are some worked examples:
| No barcodes | 12 barcodes | 24 barcodes | 24 barcodes (Flongle) |
|---|
| Flow cell price | $500 | $500 | $500 | $90 (Flongle flow cell) |
| Library price | $90 | $120 | $150 | $150 |
| Price per sample | $590 | $51.67 | $27.08 | $10 |
The 16S Barcoding Kit 24 V14 offers a method of amplifying the entire ~1,500 bp 16S rRNA gene from extracted gDNA, followed by library preparation. 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. On the other hand, analysing the full 16s rRNA gene is more informative than analysing a subset of exons.
The 16S Barcoding Kit 24 V14 features:
| Feature | Property |
|---|
| Preparation time | 25 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 14 |
| Associated protocols | Rapid sequencing DNA - 16S Barcoding Kit 24 V14 (SQK-16S114.24) |
| Multiplexing options | • 16S Barcoding Kit 24 V14
• 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 |
The DNA is amplified by PCR using specific 16S primers (27F and 1492R) that contain 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters.
This kit has been upgraded to use our Kit 14 chemistry, which includes improved modal raw read sequencing accuracies with high output on our latest nanopore: R10.4.1. The flow cell priming and sequencing reagents are also included as part of the sequencing kit and have been reformulated to be compatible with the Kit 14 chemistry upgrade and R10.4.1 nanopore. Other updates include higher capture rate of gDNA 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.
The kit is supported by the EPI2ME wf-16S workflow, which can be used to analyse data from the 16S Barcoding Kit 24 V14 (SQK-16S114.24) protocol. Deconvolution of barcoded sequencing data is supported by Dorado 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 5’ tags which facilitate the ligase-free attachment of Rapid Sequencing Adapters. The kit contains 24x barcoded primer pairs.
The kit is supported by the EPI2ME wf-16S workflow, that can be used to analyse data from the 16S Barcoding Kit 24 V14 (SQK-16S114.24) protocol. Deconvolution of barcoded sequencing data is supported by Dorado and EPI2ME, which classify the barcode sequence and sort reads into corresponding folders.
What's in the box
The 16S Barcoding Kit 24 V14 contains 24 unique barcodes and sufficient reagents to generate six sequencing libraries:
| Name | Acronym | Cap colour | No. of vials | Fill volume per vial (μl) |
|---|
| 16S Barcode Primers 01-24 | 1-24 | - | 2 plates, 3 sets of barcodes per plate | 15 μl per well |
| Rapid Adapter | RA | Green | 1 | 15 |
| Adapter Buffer | ADB | Clear | 1 | 100 |
| AMPure XP Beads | AXP | Clear cap, light teal label | 1 | 6,000 |
| Elution buffer | EB | Black | 1 | 1,500 |
| EDTA | EDTA | Blue | 1 | 700 |
| Sequencing Buffer | SB | Red | 1 | 700 |
| Library Beads | LIB | Pink | 1 | 600 |
| Library Solution | LIS | White cap, pink label | 1 | 600 |
| Flow Cell Flush | FCF | Clear cap, light blue label | 1 | 8,000 |
| Flow Cell Tether | FCT | Purple | 1 | 200 |
Note: This product contains AMPure XP Reagent Manufactured by Beckman Coulter, Inc. and can be stored at -20°C with the kit without detriment to reagent stability.