Real-Time PCR

The real-time polymerase chain reaction (PCR) uses fluorescent reporter molecules to monitor the production of amplification products during each cycle of the PCR reaction. This combines the DNA amplification and detection steps into one homogeneous assay and obviates the need for gel electrophoresis to detect amplification products. Appropriate data analysis and/or use of apposite chemistries also eliminates the need for Southern blotting or DNA sequencing for amplicon identification. Its simplicity, specificity, and sensitivity, together with its potential for high throughput and the ongoing introduction of new chemistries, more reliable instrumentation, and improved protocols, has made realtime PCR the benchmark technology for the detection of DNA.