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The bioluminescent RNA assay is a nucleic acid detection technique, which is based on the detection of released inorganic pyrophosphate (PPi) during oligonucleotide synthesis by the polymerase extension reaction. The target nucleic acid molecule can be either RNA or DNA. In a series of three consecutive enzymatic reactions the visible light is generated, which is directly proportional to the number of incorporated nucleotides. During the nucleic acid polymerization reaction the inorganic pyrophosphate PPi is released as the result of nucleotide incorporation by polymerase. The released PPi is converted into ATP by ATP-sulfurylase, which, subsequently, provides the energy for luciferase to generate light. The PPi measurement technique is extremely sensitive and can be used to quantify number of target RNA molecules and monitor RNA transcription in real time. |
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Replication of RNA and DNA produces one inorganic pyrophosphate
(PPi) molecule per each incorporated dNTP. A luminescent PPi assay detects, in
real-time, the rate of dNTP incorporation and can be used to
measure the number of mRNA targets and kinetics parameters of reverse
transcription (RT) reaction. |
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Y.
Sun, K.B. Jacobson, V. Golovlev, “A multienzyme bioluminescent
time-resolved pyrophosphate assay.”, Anal Biochem. 2007 Aug 15;367(2):201-9. |
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Sun Y., Jacobson BK.,
Golovlev V., “Quantitative Bioluminescent RNA Assay”, in Bioluminescence and Chemoluminescence: Chemistry, Biology and
Applications, Editors A. Szalay, P. Hill, L. Kricka, and P. Stanley,
World Scientific Publishing Co., 2007. |
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Sci-Tec, Inc. 2007 |
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