
doi: 10.2144/98254st01
pmid: 9793641
Methods for detecting and measuring the success of nucleic acid sequence amplifications can be developed by detecting the by-products of amplification procedures. One method includes the detection of inorganic phosphate (Pi) during or on completion of the PCR. The method requires modification of assay conditions to prevent thermal- and template-independent enzymatic activity from nonspecifically hydrolyzing dNTPs. Detection of Pi by the traditional Fiske-SubbaRow method provides a sensitivity similar to ethidium bromide staining of amplified products. The method offers a simple and rapid assay for amplified nucleic acids and can be useful in assays where confirmation of the amplified DNA product is not essential.
Molybdenum, QH301-705.5, Hydrolysis, DNA, Deoxycytidine Monophosphate, Templates, Genetic, Hydrogen-Ion Concentration, Polymerase Chain Reaction, Sensitivity and Specificity, Phosphates, Immunoglobulin Fab Fragments, Deoxyadenine Nucleotides, Spectrophotometry, Ethidium, Animals, Biology (General), Pyrophosphatases
Molybdenum, QH301-705.5, Hydrolysis, DNA, Deoxycytidine Monophosphate, Templates, Genetic, Hydrogen-Ion Concentration, Polymerase Chain Reaction, Sensitivity and Specificity, Phosphates, Immunoglobulin Fab Fragments, Deoxyadenine Nucleotides, Spectrophotometry, Ethidium, Animals, Biology (General), Pyrophosphatases
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