Molecular beacons with JOE dye: Influence of linker and 3′ couple quencher
| Autor: | Yury V. Martynenko, Vadim V. Shmanai, Maksim V. Kvach, Timofei S. Zatsepin, Sergey K. Zavriev, Andrey A. Formanovsky, Alexander A. Stakheev, Ilya O. Aparin, Sergey V. Gontarev, Dmitry Y. Ryazantsev, Igor A. Prokhorenko, Dmitry A. Tsybulsky, Vladimir A. Korshun |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Dna template Stem length Biology Nucleic Acid Denaturation 010402 general chemistry Polymerase Chain Reaction 01 natural sciences Fusarium avenaceum 03 medical and health sciences chemistry.chemical_compound Molecular beacon Fluorescein Molecular Biology Fluorescent Dyes Base Sequence Temperature Cell Biology Reference Standards Fluoresceins Fluorescence 0104 chemical sciences Elongation factor 030104 developmental biology Xanthenes chemistry Molecular Probes Biophysics DNA Probes Linker |
| Zdroj: | Molecular and Cellular Probes. 30:285-290 |
| ISSN: | 0890-8508 |
| Popis: | Molecular beacons carrying JOE dye (4',5'-dichloro-2',7'-dimethoxy-6-carboxyfluorescein) on a rigid or flexible linker and one or two BHQ1 quenchers have been prepared and tested in real-time PCR using Fusarium avenaceum elongation factor 1α DNA template. The probes were different in their structures (loop size and stem length), linkers for dye attachment (6-aminohexanol or trans-4-aminocyclohexanol), quencher composition (single and double BHQ1) to elucidate the influence of all these features. Fluorogenic properties of the probes were studied and compared to those of FAM (fluorescein)-based probes. All the factors - stem length, JOE vs FAM, rigid vs flexible linker, single vs double quencher - appeared to play a considerable role in the probe's fluorescent properties and determine the usability of the probe at two different temperatures of fluorescence detection (55°С and 64°С). |
| Databáze: | OpenAIRE |
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