Dual-Labeled probes are the most common probe type for qPCR and are often referred to as hydrolysis probes. These versatile probes can be used for a variety of applications including:
Dual-labeled probes are perfect for validating MISSION® siRNA and shRNA knockdowns. Their benefits include design simplicity for sequence specificity, extensive availability of reporter/quencher combinations, and increased sensitivity.
Adding locked nucleic acids to your probe increases thermal stability and hybridization specificity. They allow greater accuracy in SNP detection, allele discrimination, and in vitro quantification or detection. Locked nucleic acids also allow easier and more sensitive probe designs for problematic target sequences.
A Dual-Labeled Probe is a single-stranded oligonucleotide labeled with two different dyes. A reporter dye is located at the 5’ end and a quencher molecule located at the 3’ end. The quencher molecule inhibits the natural fluorescence emission of the reporter by fluorescence resonance energy transfer (FRET). The illustration below depicts the mechanism.
The primer is elongated by the polymerase and the probe binds to the specific DNA template. Hydrolysis releases the reporter from the probe/target hybrid, causing an increase in fluorescence. The measured fluorescence signal is directly proportional to the amount of target DNA.
Design and Customize your qPCR Probes to best suit your application.
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