Colorimetric method for detecting amplified nucleic acids.

E Durward; W J Harris

doi:10.2144/98254st01

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PubMed · 9793641

Colorimetric method for detecting amplified nucleic acids.

E Durward·W J Harris

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Abstract

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.

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Animals DNA Deoxyadenine Nucleotides Deoxycytidine Monophosphate Ethidium Hydrogen-Ion Concentration Hydrolysis Immunoglobulin Fab Fragments Molybdenum Phosphates Polymerase Chain Reaction Pyrophosphatases Sensitivity and Specificity Spectrophotometry Templates, Genetic Technical Report Journal Article

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E Durward, W J Harris. 1998. Colorimetric method for detecting amplified nucleic acids.. https://doi.org/10.2144/98254st01

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