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Mark Aaron Behlke Integrated DNA Technologies, Inc., United States of America |
Abstract
Fluorescence-quenched probes are used in a wide variety of molecular biology applications today and have particular utility in qPCR. Efficient quenching is crucial for optimal probe performance. A new dark quencher (ZEN) has been developed which can be placed internally within hydrolysis probes, permitting the quencher to be positioned very close to the fluorophore, thereby improving quenching and probe performance. Most chemical groups are destabilizing to duplex formation when introduced internally within a sequence; the new ZEN quencher stabilizes duplex formation and thus increases Tm of the probe instead of lowering Tm as would normally be expected.
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