The Illumina Sequencing Protocol and the NovaSeq 6000 System

Alessandra Modi; Stefania Vai; David Caramelli; Martina Lari

doi:10.1007/978-1-0716-1099-2_2

The Illumina Sequencing Protocol and the NovaSeq 6000 System

Methods Mol Biol. 2021:2242:15-42. doi: 10.1007/978-1-0716-1099-2_2.

Authors

Alessandra Modi¹, Stefania Vai¹, David Caramelli¹, Martina Lari²

Affiliations

¹ Department of Biology, University of Firenze, Firenze, Italy.
² Department of Biology, University of Firenze, Firenze, Italy. martina.lari@unifi.it.

PMID: 33961215
DOI: 10.1007/978-1-0716-1099-2_2

Abstract

The NovaSeq 6000 is a sequencing platform from Illumina that enables the sequencing of short reads with an output up to 6 Tb. The NovaSeq 6000 uses the typical Illumina sequencing workflow based on library preparation, cluster generation by in situ amplification, and sequencing by synthesis. Flexibility is one of the major features of the NovaSeq 6000. Several types of sequencing kits coupled with dual flow cell mode enable high scalability of sequencing outputs to match a wide range of applications from complete genome sequencing to metagenomics analysis. In this chapter, after explaining how to assemble a normalized pool of libraries for sequencing, we will describe the experimental steps required to run the pools on the NovaSeq 6000 platform.

Keywords: Library quality control; Library quantification; Run setup; Sequencing pool; Standard sequencing workflow; XP sequencing workflow.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Genomic Library
Genomics* / instrumentation
High-Throughput Nucleotide Sequencing* / instrumentation
Research Design
Sequence Analysis, DNA* / instrumentation
Workflow