Open AccessColorimetric Method for Detecting Amplified Nucleic Acids
Author(s) -
Elaine Durward,
William J. Harris
Publication year - 1998
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/98254st01
Subject(s) - nucleic acid , ethidium bromide , nucleic acid quantitation , dna , microbiology and biotechnology , chemistry , biochemistry , nucleic acid methods , chromatography , biology
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.