Rapid Sequencing Kits offer a simple and rapid library preparation method for genomic DNA sequencing on MinION™ devices.

• Sequence native DNA — eliminate PCR bias and retain base modifications
• Fast library prep — in as little as 10 minutes from extracted DNA
• Minimal lab equipment required
• Multiplex samples with rapid barcoding kits

Rapid Sequencing Kits generate sequencing libraries from extracted gDNA in as little as 10 minutes* using a simple, streamlined, workflow — requiring minimal lab equipment. With no requirement for amplification, these kits eliminate potential PCR bias and preserve base modifications (e.g. methylation), which can be detected alongside the nucleotide sequence.

These kits use a transposase to simultaneously cleave template molecules and attach tags to the cleaved ends. Rapid Sequencing Adapters are then added to the tagged ends.

Due to the simple nature of the workflow and the fact that little sample manipulation is required (e.g. minimal pipetting steps and no clean-up steps), some very long reads can be achieved with these kits, despite the required transposase fragmentation. However, in order for long reads to be observed in sequencing, long DNA fragments must be present in the sample.

In addition to the singleplex Rapid Sequencing Kit, which enables one sample to be sequenced per run, two standalone rapid barcoding kits are available: the Rapid Barcoding Kit 24 and Rapid Barcoding Kit 96, which enable multiplexing of up to 12 and 96 samples per run, respectively. 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.

Rapid Sequencing and Barcoding Kits V14 ― the latest nanopore sequencing chemistry, which uses R10.4.1 Flow Cells to deliver improved median raw read sequencing accuracies and higher output. The flow cell priming and sequencing reagents are now included as part of the sequencing kit and have been reformulated to be compatible with the chemistry upgrade and R10.4.1 Flow Cells. These kits are able to generate Q20+ and duplex data but have been optimised for speed and simplicity. If users require high duplex data, we recommend using the Ligation Sequencing Kit V14. The Rapid Sequencing Kit V14 recommends a total input of 100 ng high molecular weight (>30 kb) gDNA per sample, while the Rapid Barcoding Kits V14 recommend 50 ng high molecular weight (>30 kb) gDNA per sample. Addition of less input DNA, or shorter fragments could compromise sequencing throughput and read length.

Rapid Barcoding Kits attach barcoded tags to the cleaved ends of the template molecules. Barcoded samples are pooled, and Rapid Sequencing Adapters are then added to the tagged ends. Deconvolution of barcoded sequencing data is supported by the MinKNOW software provided with Oxford Nanopore sequencing devices, which classify the barcode sequence and sort reads into corresponding folders.

*Rapid Sequencing Kit V14. Approximately 60 minutes for Rapid Barcoding Kits.