Sequencing technology:
Samples collection and analysis, an article.
Sangar technique basis:
Pyrosequencing -
454 developed an array-based pyrosequencing technology which has emerged as a platform for large-scale DNA sequencing. Roche announced the discontinuation of the 454 sequencing platform in 2013.
two approches:
a) sequencing by synthesis:
A DNA segment is amplified and the strand to serve as the Pyrosequencing template is biotinylated. After denaturation, the biotinylated single-stranded PCR amplicon is isolated and allowed to hybridize with a sequencing primer. Each incorporation event is accompanied by the release of pyrophosphate (PPi) in a quantity equimolar to the amount of incorporated nucleotide. ATP sulfurylase converts PPi to ATP in the presence of adenosine 5' phosphosulfate (APS). This ATP drives the luciferase-mediated conversion of luciferin to oxyluciferin that generates visible light in amounts that are proportional to the amount of ATP.
For more information:
Large-scale DNA sequencing techniques