Roche

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Roche 454 DNA Sequencing Images adapted from
Teri Rambo Mueller Roche Diagnostics Corporation
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Wells are picoliter reaction vessels. Wells
have titanium coating. Each well holds 1 bead.
Multiple optical fibers fused in array.
Light-producing reactions in wells measured by
CCD camera.
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oxy luciferin light
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Polymerase adds nucleotide Pyrophosphate (PPi)
released Sulfurylase PPi ? ATP Luciferase
ATP luciferin ? light
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Repeated dNTP flow sequence
G
C
T
A
Process continues until user-defined number of
nucleotide flow cycles are completed.
A A T C G G C A T
G C T A A A A C T G
A
A
A
G
C
T
A
T
T
T
G
C
C
G
C
G
A
C
G
T
T
T
T
T
A
Anneal Primer
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• Polymerase adds nucleotide (dATP)
• Pyrophosphate is released (PPi)
• Sulfurylase creates ATP from PPi
• Luciferase hydrolyses ATP and uses luciferin to
  make light

DNA Bead with clonally amplified fragments