Molecular beacons with JOE dye: Influence of linker and 3 ' couple quencher

作者：Tsybulsky Dmitry A; Kvach Maksim V; Ryazantsev Dmitry Y; Aparin Ilya O^*; Stakheev Alexander A; Prokhorenko Igor A; Martynenko Yury V; Gontarev Sergey V; Formanovsky Andrey A; Zatsepin Timofei S; Shmanai Vadim V; Korshun Vladimir A; Zavriev Sergey K^*

来源：Molecular and Cellular Probes, 2016, 30(5): 285-290.

DOI：10.1016/j.mcp.2016.10.003

• Real-time PCR
• Molecular beacons
• JOE dye
• Linkers
• Couple quencher

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 1 alpha 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 degrees C and 64 degrees C).

• 出版日期2016-10