Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
2006-11-28
2006-11-28
Strzelecka, Teresa E. (Department: 1637)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C435S091200
Reexamination Certificate
active
07141370
ABSTRACT:
The present invention concerns methods of quantifying nucleic acids using a bioluminescence regenerative cycle (BRC). In BRC, steady state levels of bioluminescence result from processes that produce pyrophosphate. Pyrophosphate reacts with APS in the presence of ATP sulfurylase to produce ATP. The ATP reacts with luciferin in a luciferase-catalyzed reaction, producing light and regenerating pyrophosphate. The pyrophosphate is recycled to produce ATP and the regenerative cycle continues. Because the kinetic properties of ATP sulfurylase are much faster than luciferase, a steady state results wherein concentrations of ATP and pyrophosphate and the rate of light production remain relatively constant. Photons are counted over a time interval to determine the number of target molecules present in the initial sample. The BRC process has a controllable dynamic range up to seven orders of magnitude and is sensitive enough to detect a few thousand molecules of target nucleic acid.
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Hassibi Arjang
Pourmand Nader
Aston David J.
Peters Verny Jones Schmitt & Aston, LLP
Strzelecka Teresa E.
The Board of Trustees of the Leland Stanford Junior University
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