Qiagen PyroMark Q96 ID User Manual page 20

General Description
4. Each incorporation event is accompanied by release of
5. ATP sulfurylase quantitatively converts PPi to ATP in the
6. This drives the conversion of luciferin to oxyluciferin by
7. Apyrase, a nucleotide-degrading enzyme, continuously
8. Nucleotides are added one at a time.
3-2
pyrophosphate (PPi) in an equimolar quantity to the
amount of nucleotide incorporated.
presence of adenosine 5' phosphosulfate.
luciferase, generating visible light in amounts
proportional to the amount of ATP. Light is detected
using charged coupled devices (CCDs) and seen as a
peak in the Pyrogram
to the number of nucleotides incorporated.
degrades unincorporated nucleotides and ATP. When
degradation is complete, another nucleotide is added.
Note: Deoxyadenosine alfa-thio triphosphate (dATPαS)
is used instead of natural deoxyadenosine triphosphate
(dATP) since it is used efficiently by the DNA polymerase,
but not recognized by the luciferase.
PyroMark Q96 ID User Manual 01/2016
® . Each light signal is proportional