Real-time PCR
Real-time PCR is a second generation PCR platform with significantly improved testing characteristics. Introduced in 1996, it has revolutionized and replaced conventional PCR approaches to quantify DNA and RNA. Today, RT-PCR is the gold standard for quantitative PCR and is rapidly becoming accepted as the method of choice for PCR diagnostics.
Contamination Control
Quantification is only one of many advantages of RT-PCR. In contrast to conventional PCR, reaction tubes stay closed after the PCR process is finished. PCR product accumulation is measured in real-time during the amplification process with a fluorescently labeled probe within the PCR reaction tube. The closed-tube detection system reduces PCR product carry-over and risk of false positive signal generation. We also use a second system in our mastermixes which does not allow PCR products to be re-amplified.
Increased Analytical Sensitivity and Specificity
Real-time has shown to be ultra-sensitive; direct comparisons with nested conventional PCR puts it ahead of the conventional approach. Real-time PCR has a lower limit of detection of 5 molecules and has better analytical sensitivity than conventional PCR with eliminated risk of contamination.
For more information about Real-time PCR please refer to the following article. “The Real-time TaqMan PCR and Applications in Veterinary Medicine.” (PDF)