Real-Time or Kinetic PCR

The DNA Facility houses the “real-time” or kinetic PCR instrument, the Applied Biosystems Model 7700 sequence detection system (the TaqMan instrument). The polymerase chain reaction (PCR) has revolutionized the detection of DNA and RNA. As little as a single copy of a particular sequence can be specifically amplified and detected. Theoretically, there is a quantitative relationship between amount of starting target sequence and amount of PCR product at any given cycle. In practice, though, it is a common experience for replicate reactions to yield different amounts of PCR product. The development of real-time quantitative PCR has eliminated the variability traditionally associated with quantitative PCR, thus allowing the routine and reliable quantification of PCR products. This instrument, therefore, now provides investigators with the ability to perform very sensitive, accurate, and reproducible measurements of levels of gene expression. In addition, this instrument can be used in other applications such as measuring viral load, performing allelic discrimination studies, and optimizing PCR conditions.