Table 1.

Summary of available ONT library preparation kits for sequencing of microbial communities.

ONT library preparation strategy	Input ng recommendation	Preparation time	Multiplexing	Application
16S Rapid Barcoding Kit	< 10 ng gDNA	10 min + PCR	Up to 12 or 24 samples	Targeted 16S rRNA gene sequencing
Rapid Sequencing Kit	≥ 400 ng HMW DNA	10 min	Up to 12 samples	Metagenomics and epigenomics, amplification-free
Rapid PCR Sequencing Kit	≤ 10 ng gDNA	15 min + PCR	Up to 12 samples	Metagenomics, requires amplification
Ligation Sequencing Kit	≥ 1000 ng dsDNA	60 min	Up to 96 samples	Metagenomics and epigenomics, amplification-free, high-throughput
PCR Sequencing Kit	≤ 100 ng gDNA	60 min + PCR	Up to 12 samples	Metagenomics, requires amplification, high-throughput
Direct cDNA Sequencing Kit	100 ng poly-A+ RNA	270 min	Up to 24 samples	Metatrascriptomics, requires retrotranscription
PCR cDNA Sequencing Kit	1 ng poly-A+ or 50 ng total RNA	165 min	Up to 12 samples	Metatranscriptomics, requires retrotranscription and amplification
Direct RNA Sequencing Kit	500 ng poly-A+ RNA	105 min	None	Metatrascriptomics and epitranscriptomics, retrotranscription- and amplification-free

HMW: high-molecular weight.