Small-Scale Illumina Library Preparation Using the Illumina Nextera XT DNA Library Preparation Kit.
Merve S ZedenAngelika GründlingPublished in: Cold Spring Harbor protocols (2023)
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.
Keyphrases
- circulating tumor
- high throughput sequencing
- cell free
- single molecule
- staphylococcus aureus
- randomized controlled trial
- quality control
- nucleic acid
- molecularly imprinted
- circulating tumor cells
- dna methylation
- pseudomonas aeruginosa
- biofilm formation
- escherichia coli
- minimally invasive
- cystic fibrosis
- mass spectrometry
- candida albicans
- study protocol