16S Barcoding Kit

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.

 

Key features:

Prep time

PCR + 10 minutes

Input amount

10 ng gDNA

Read length

= full-length 16S gene (~1.5kb)

Typical throughput

1-2 Gb in 6 hours, 4-8 Gb in 48 hours per flow cell on MinION/GridION

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 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 Albacore and EPI2ME which classify the barcode sequence and sort reads into corresponding folders.