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
• want to optimise their sequencing experiment for throughput
• require control over read length
• are interested in utilising upstream processes such as size selection or whole genome amplification.

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.

Workflow

The library preparation method is similar to the Ligation Sequencing Kit protocol; DNA ends are repaired and dA-tailed using the NEBNext End Repair/dA-tailing module, and then a unique dT-tailed barcode adapter is ligated on the dA-tailed template. Barcoded samples are then pooled together. Each barcode adapter also has a cohesive end and this is used as a hook to ligate to the supplied sequencing adapters.