Real-time PCR is a fast developing field with wide ranging applications for researchers. From gene expression analysis from cDNA, population genotyping studies, to the multiplex detection of several disease targets real-time PCR has opened the door to a world of possibilities.
Real-time analysis is broadly divided into two areas; probe and intercalating dye based assays. The two techniques have many overlapping applications.
For the majority of applications we recommend the use of an intercalating dye technology rather than costly dual labelled probes. Using an intercalating dye, such as SyGreen, rather than a hydrolysis probe (such as Taqman®) has the added benefit of post amplification melt profile analysis. Intercalating dye real-time PCR also has the advantage of being able to cycle faster than with hydrolysis probes, this is due to the slower rate of 5′ to 3′ exonuclease activity of taq DNA polymerase compared to its polymerisation rate.
The principle advantage of probe based technologies is that multiple amplicons can be detected in the same tube. The number of amplicons that can be detected in real-time PCR is typically limited by the number of light channels found in the real-time PCR instrument. PCR Biosystems probe mixes are designed not to lose efficiency during multiplexing. PCR Biosystems real-time PCR probe mixes have been designed for use on a wide range of probe technologies including Taqman®, Molecular Beacons® and Scorpion probes®.
All PCR Biosystems real-time PCR mixes can be run under fast or standard cycling conditions.