Small-Scale Illumina Library Preparation Using the Illumina Nextera XT DNA Library Preparation Kit
- 1Microbiology, School of Biological and Chemical Sciences, National University of Galway, Galway H91 TK33, Ireland
- 2Section of Molecular Microbiology and Medical Research Council Centre for Molecular Bacteriology and Infection, Imperial College London, London SW7 2AZ, United Kingdom
- ↵3Correspondence: a.grundling{at}imperial.ac.uk
Abstract
Here, we describe a protocol for a scaled-down version of a genomic DNA (gDNA)-fragmentation and tagmentation reaction using the Illumina Nextera XT DNA Library Preparation Kit. Using Staphylococcus aureus as an example, which has a genome size of ∼3 Mb, we show how 24 different samples can be pooled for a typical paired-end Illumina high-throughput sequencing run using the MiSeq Reagent V2 300-cycle kit, with which it is possible to sequence 5.1 Gb of DNA. As part of the protocol, a DNA size-selection method using a standard DNA agarose gel–extraction procedure and a final sample quality-control step using a Bioanalyzer are described.
Footnotes
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From the Experiments in Bacterial Genetics collection, edited by Lionello Bossi, Andrew Camilli, and Angelika Gründling.
