Nucleic acid amplification with terminal-phosphate labeled...

Details Nucleic acid amplification with terminal-phosphate labeled... Nucleic acid amplification with terminal-phosphate labeled...

2006-10-24

2006-10-24

Horlick, Kenneth R. (Department: 1637)

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

C435S091100, C435S091200

Reexamination Certificate

active

07125671

ABSTRACT:
The present invention relates generally to the use of terminal-phosphate-labeled nucleotides having three or more phosphates as substrates for nucleic acid polymerases and their use in DNA amplification. The labels employed are chemiluminescent, fluorescent, electrochemical and chromogenic moieties as well as mass tags and include those that are directly detectable, detectable after enzyme activation or feed into other processes to generate a different signal. The signal generated from the attached dyes may also be used to quantify the amount of amplification. Further provided are stabilizers that enhance the stability of terminal-phosphate labeled nucleoside polyphosphates in aqueous solutions and are useful for reducing non-enzymatic hydrolysis of these nucleotides, hence decrease background.

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Dyatkina, N., et al., “Modified Triphosphates of Carbocyclic Nucleoside Analogues: Synthesis, Stability Towards Alkaline Phosphatase and Substrate Properties for some DNA Polymerases”,Bioorganic&Medicinal Chemistry Letters,vol. 6, No. 22, 1996, p. 2639-2642.
Vassiliou, W., et al., “Exploiting Polymerase Promiscuity: A Simple Colorimetric RNA Polymerase Assay”,Virology,vol. 274, 2000, p. 429-437.

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