Terminal-phosphate-labeled nucleotides and methods of use

A method of detecting the presence of a DNA sequence comprising the steps of:

• a) conducting a DNA polymerase reaction in the presence of a terminal-phosphate-labeled nucleotide, which reaction results in the production of a labeled polyphosphate;
• b) permitting said labeled polyphosphate to react with a phosphatase to produce a detectable species;
• and
• c) detecting the presence of said detectable species.

A method of quantifying a nucleic acid comprising the steps of:

• (a) conducting a nucleic acid polymerase reaction, said reaction comprising the reaction of nucleotide which is substantially non-reactive to phosphatase and at least one terminal-phosphate-labeled nucleotide;
• said reaction resulting in production of labeled polyphosphate;
• (b) permitting said labeled polyphosphate to react with a phosphatase to produce a detectable by-product species in amount substantially proportional to the amount of said nucleic acid;
• (c) measuring said detectable species;
• and
• (d) comparing said measurements using known standards to determine the quantity of nucleic acid.

A method of quantifying a DNA sequence comprising the steps of:

• (a) conducting a DNA polymerase reaction in the presence of a terminal-phosphate-labeled nucleotide, said reaction resulting in production of labeled polyphosphate;
• (b) permitting said labeled polyphosphate to react with a phosphatase to produce a detectable by-product species in amounts substantially proportioned to the amount of said DNA sequence;
• (c) measuring said detectable species;
• and
• (d) comparing said measurements using known standards to determine the quantity of DNA.

A method of detecting the presence of a nucleic acid sequence comprising the steps of:

• (a) conducting a nucleic acid polymerase reaction in the presence of at least one terminal-phosphate-labeled nucleotide having four or more phosphate groups in the polyphosphate chain, which reaction results in the production of a labeled polyphosphate;
• (b) permitting said labeled polyphosphate to react with a phosphatase to produce a detectable species;
• and
• (c) detecting the presence of said detectable species.