1) Nebuliser cuts DNA into 300-800 fragments.
2) Lengths degraded to single stranded DNA. (TEMPLATES)
3) Incubated with; primer, DNA polymerase, ATP sulfurylase, luciferase, apyrase, APS, luciferin and one of 4 activated nucleotides.
4) When activated nucleotide incorporated;
- 2 phosphoryls released
- in APS presence, phosphoryls converted to ATP
- in presence of ATP, luciferase (enzyme) converts luciferin to oxyluciferin.
- conversion generates light; detected via camera.
Amount of light generated = proportional to amount of ATP released (depends on diff nucleotides)
Unincorporated nucleotides broken down via Apyrase.
Software will package the strands into longer sequences.
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