Nanopore Sequencing Kit Protocols

These protocols use information from Oxford Nanopore Technologies' protocols and have been written by me; I release them to the world under a CC-BY-SA license. The source .Rmd text for all versions can be made available; please let me (David Eccles) know if that would be useful to you.

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PCR cDNA Barcoding Kit Protocol

The PCR cDNA Kit is used for high-accuracy quantification of cDNA from extracted RNA. If sample amounts are abundant, the number of cDNA cycles can be reduced in order to lessen the impact of PCR-related artefacts.

Rapid Barcoding Kit Protocol

The Rapid Barcoding Kit is a cheap, fast protocol; great for plasmids, amplicons, and any situations where there is lots of high-quality DNA. Because the transposase fragments DNA, it can be used on circular DNA (e.g. plasmids, mitochondria) without any additional linearisation.

Ligation Sequencing Kit Protocol

The Ligation Sequencing Kit is a stable / robust protocol that delivers excellent sequencing yields; great for large genome sequencing, or situations where getting the highest yield from a sample is important. Note that because this is a ligation protocol, there is a higher chance of ligation-associated artefacts (chimeric reads) being generated during the sample preparation process.

Rapid PCR Barcoding Kit Protocol

The Rapid PCR Barcoding Kit is a moderately cheap and fast protocol that works great for metagenomic applications, and any situations where sample amounts are low (less than 5ng). It produces read lengths of about 1-3kb, which is usually fine for microbial assembly, but can be too short for large genome assembly