Fluorescent Primer(s) Creation for Nucleic Acid Detection and Amplification

CDC researchers have developed technology that consists of a simple and inexpensive technique for creating fluorescent labeled primers for nucleic acid amplification. Fluorescent chemical-labeled probes and primers are extensively used in clinical and research laboratories for rapid, real-time detection and identification of microbes and genetic sequences. During nucleic acid amplification, the "UniFluor" primer is incorporated into newly synthesized double stranded DNA. As a consequence, quenching of the dye's fluorescent signal occurs decreasing the fluorescence of the sample several fold. The decrease in fluorescence can be measured and observed using any commercially available nucleic acid amplification system that measures fluorescence (e.g., real-time PCR/thermocyclers). Because many real-time PCR applications require a multitude of fluorescently labeled primers or probes, the single-labeled primer technique also allows researchers and clinicians to perform their work at lower cost.