Fig. 1
From: Some considerations on the current debate about typing resolution in solid organ transplantation
An overview of DNA sequencing by synthesis technology (Illumina). After random fragmentation of genomic or cDNA, adapters, which are specialized oligonucleotides, are bound to both 5′ and 3′ DNA fragment ends, allowing ligation and NGS library preparation. The library is loaded into a flow cell that is a glass slide with one (for MiSeq technology) or eight lanes (for HiSeq, allowing eight independent experiments) coated with surface-bound, adapter-complimentary oligos. The sample is hybridized onto the flow cell, generating a cluster and is clonally amplified through bridge amplification until the cluster has 1000 copies. Each cluster on the flow cell produces a single sequence read. The flow cell is imaged, and the emission from each image is recorded. Finally, the reads are aligned to a reference sequence using bioinformatics tools