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A novel FRET pair for detection of parallel DNA triplexes by the LightCycler |
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| Authors: | Uffe V Schneider Jette K Severinsen Imrich Géci Limei M Okkels Nina Jøhnk Nikolaj D Mikkelsen Teena Klinge Erik B Pedersen Henrik Westh Gorm Lisby |
| Affiliation: | (1) QuantiBact Inc, Department of Clinical Microbiology, Hvidovre Hospital, Kettegaards Alle 30, 2650 Hvidovre, Denmark;(2) Nucleic Acid Center and Department of Chemistry, University of Southern Denmark, Campusvej 55, 5230 Odense M, Denmark;(3) Department of Clinical Microbiology, Hvidovre Hospital, Kettegaards Alle 30, 2650 Hvidovre, Denmark;(4) Faculty of Health, University of Copenhagen, Copenhagen, Denmark |
| Abstract: | BackgroundMelting temperature of DNA structures can be determined on the LightCycler using quenching of FAM. This method is very suitable for pH independent melting point (Tm) determination performed at basic or neutral pH, as a high throughput alternative to UV absorbance measurements. At acidic pH quenching of FAM is not very suitable, since the fluorescence of FAM is strongly pH dependent and drops with acidic pH. |
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