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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
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 | |
---|---|---|---|
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 |
PCR Required | Yes |
Fragmentation | Not required |
Read length | Full-length 16S gene (~1.5 kb) |
Read type produced | 1D |
Typical throughput* | ••• |
Associated protocols |
16S Barcoding Kit Flow Cell Wash Kit |
Pack size | 6 reactions |
Stability |
Shipped at 2-8° C Long-term storage -20° C |
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.
Further considerations:
Oxford Nanopore Technologies deem the useful life of the product to be 3 months from receipt by the customer.