Quantitative PCR Technology

Autor: Susan J. A. Flood, John Shigeura, Dane Carlson, Mark F. Oldham, Jeff Lucas, Traci Allen, Peter Honebein, Kevin S. Bodner, Linda Lee, Ken Livak, Raymond Lefebvre, Daniel Thiel, Robert Grossman, Lincoln J. Mcbride, Bashar Mullah, Mike Lucero, Bruce Goldman, Junko Stevens, Federico Goodsaid, Eugene Young, Charles Connel, Tim Woudenberg, Paul Wyatt
Rok vydání: 1998
Předmět:
Zdroj: Gene Quantification ISBN: 9781461286820
DOI: 10.1007/978-1-4612-4164-5_6
Popis: Higuchi et al. (1992, 1993) pioneered the analysis of PCR kinetics by constructing a system that detects PCR products as they accumulate. This “real-time” system includes the intercalator ethidium bromide in each amplification reaction, an adapted thermal cycler to irradiate the samples with ultraviolet light, and detection of the resulting fluorescence with a computer-controlled cooled CCD camera. Amplification produces increasing amounts of double-stranded DNA, which binds ethidium bromide, resulting in an increase in fluorescence. By plotting the increase in fluorescence versus cycle number, the system produces amplification plots that provide a more complete picture of the PCR process than assaying product accumulation after a fixed number of cycles.
Databáze: OpenAIRE