Advantages of Real-time PCR over traditional PCR
Direct visualization of the amplification process
The Real-time PCR system is based on the direct detection and quantitation of a Reporter fluorescent signal that increases proportionally to the number of amplicons generated. Data can therefore be collected at any point of the amplification. Traditional PCR only allows end-point detection.
Precise quantitation possible
Real-time PCR allows quantifying DNA molecules over an extraordinarily wide range (at least 5 log units) – Referemce #1. This technique can detect as little as a two-fold change in yield. In comparison, the agarose gel resolution only allows detection of a minimum of five to ten-fold changes in yield.
Similar or higher sensitivity
Real-time PCR allows the detection of very minute amounts of the targeted DNA sequence (less than five copies of a target DNA molecule in some cases (1).
No post-PCR processing
Traditional PCR requires the use of agarose gel to visualize and analyze the final PCR product. Real-time PCR does not require any post-PCR manipulations, thereby greatly decreasing the analytic turnaround time of each reaction while minimizing the chances for cross contamination in the laboratory (1).
- Mark A. Valasek1 and Joyce J. Repa, The power of real-time PCR; Adv Physiol Educ 29: 151–159, 2005.
- Reischl U, Wittwer CT, and Cockerill F. Rapid Cycle Real-time PCR: Methods and Applications; Microbiology and Food Analysis. New York:Springer-Verlag, 2002.