Small-Scale Illumina Library Preparation Using the Illumina Nextera XT DNA Library Preparation Kit
- 28 April 2023
- journal article
- Published by Cold Spring Harbor Laboratory in Cold Spring Harbor Protocols
- Vol. 2023 (11)
- https://doi.org/10.1101/pdb.prot107900
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.Keywords
This publication has 2 references indexed in Scilit:
- Preparation ofStaphylococcus aureusGenomic DNA Using Promega Nuclei Lysis and Protein Precipitation Solutions, Followed by Additional Cleanup and Quantification StepsCold Spring Harbor Protocols, 2023
- Preparation ofStaphylococcus aureusGenomic DNA Using a Chloroform Extraction and Ethanol Precipitation Method, Followed by Additional Cleanup and Quantification StepsCold Spring Harbor Protocols, 2023