Category:ECO:0000223 ! 454 pyrosequencing evidence

name: 454 pyrosequencing evidence
def: "A type of high throughput nucleotide sequencing evidence in which the sequence for a ssDNA is determined by addition of primers and then nucleotides, one base pair at a time (which generate a specific fluorescence), to the target strand." [EFO:0005016]
synonym: "pyrosequencing" BROAD [OBI:0000730]
is_a: ECO:0000221 ! high throughput nucleotide sequencing assay evidence
comment: 454 pyrosequencing technology amplifies individual DNA fragments inside water droplets within an oil solution (emulsion PCR). Within each droplet, a single DNA template is attached to a single primer-coated bead. Each bead is subsequently captured in a picolitre well within the sequencing machine. A luciferase-based readout system is used to identify individual nucleotides added to the nascent DNA.\n\nThe target strand is usually prepared by fragmentation, adaptor ligation, and amplification by emulsion PCR.