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Double-Quenched Probes Increase PCR Sensitivity

By LabMedica International staff writers
Posted on 18 May 2010
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Tools available for researchers in molecular biology and genomics are constantly being refined and improved. This is especially true when the method of choice is qualitative real-time polymerase chain reaction (qPCR).

Real-time PCR requires the highest quality fluorescent and quencher-labeled oligonucleotide probes. Traditionally, this has meant probes with 20-30 bases between the fluorescent dye and the quencher molecule. Now, Integrated DNA Technologies (Coralville, IA, USA) has introduced a new double-quenched probe, which increases the accuracy and effectiveness of 5' nuclease qPCR experiments. This novel proprietary probe contains an internal quencher only nine bases from the 5' fluorescent label. This shortened distance, particularly when combined with the standard 3' quencher, significantly decreases background fluorescence and increases sensitivity. Since the initial background fluorescence signal is much lower than with traditional probes, it is much easier to detect subsequent changes and, therefore, functionally increases assay sensitivity.

Integrated DNA Technologies is one of the largest suppliers of custom nucleic acids in the United States, serving academic, government, and commercial researchers in biotechnology, clinical diagnostics, and pharmaceutical development.

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