Lee Kisay, Kim Kyungju
Department of Environmental Engineering and Biotechnology, Myongji University, Yongin, Kyongki, 449-728, Korea.
Biotechnol Lett. 2003 Oct;25(20):1739-42. doi: 10.1023/a:1026035628935.
A simple and quick colorimetric method for confirming DNA amplification in polymerase chain reactions (PCR) is described and has been applied to the amplification of Giardia lamblia DNA. This method detects the release of pyrophosphate based on the competition between 1, 10-phenanthroline and pyrophosphate complexing with ferrous ion. When 1, 10-phenanthroline complexed with Fe2+ is added to the finished PCR solution, depending on whether or not the DNA was amplified, the mixture is, respectively, either bleached or red. The color changed optimally for 20-30 min at 60-80 degrees C, and the result could be determined by detecting an absorbancy change at 510 nm or a color change discernible to the naked eye. The extent of change in absorbance was proportional to the amount of pyrophosphate produced.
