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#Ion Torrent sequencing

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Sequencing

Ion Torrent sequencing is also called:

Ion Torrent platform is frequently termed pH-mediated or semiconductor sequencing because its detection is based on hydrogen ions liberated during polymerization. The sensor chip contains millions of ion-sensitive field-effect transistors that detect small pH shifts rather than fluorescence. Nanopore sequencing measures ionic current blockade as DNA traverses protein pores, SBS denotes sequencing-by-synthesis with fluorescent reversible terminators used by Illumina, and tandem sequencing is not a standard term. Consequently, pH-mediated sequencing accurately describes Ion Torrent chemistry and distinguishes it from optical or nanopore methods.

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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Sequencing

Ion Torrent sequencing detects:

Ion Torrent semiconductor sequencing, introduced by Life Technologies, determines sequence without optics. Each microwell contains template-coated beads and a polymerase. When a deoxynucleotide complementary to template is incorporated, a covalent phosphodiester bond forms and a proton, hydrogen ion, is released as byproduct. This release transiently acidifies microwell solution, typically by 0.02 pH units, detected by an ion-sensitive field-effect transistor beneath well. Signal amplitude reflects homopolymer length. Detection principle is therefore pH change, not light, electrical charge of DNA itself or fluorescence employed in other platforms.

Ref: NCERT Biology Class XII Principles on Klenow fill-in labeling, Lehninger Chapter 9 DNA cloning techniques, and Molecular Cloning by Sambrook Chapter 10 documenting end-labeling of cohesive termini.

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