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  • Title: Solid- and solution-phase synthesis and application of R6G dual-labeled oligonucleotide probes.
    Author: Skoblov AY, Vichuzhanin MV, Farzan VM, Veselova OA, Konovalova TA, Podkolzin AT, Shipulin GA, Zatsepin TS.
    Journal: Bioorg Med Chem; 2015 Oct 15; 23(20):6749-56. PubMed ID: 26392371.
    Abstract:
    A novel N-TFA-protected carboxyrhodamine 6G (R6G) phosphoramidite was synthesized for use in an automated DNA synthesis to prepare 5'-labeled oligonucleotides. Deprotection and purification conditions were optimized for 5'-labeled and dual-labeled oligonucleotide probes. As an alternative we synthesized an azide derivative of R6G for CuAAC post-synthetic oligonucleotide labeling. Dual-labeled probes obtained by both methods showed the same efficacy in a quantitative PCR assay. R6G-labeled probes demonstrated superior properties in a qPCR assay in comparison with alternative HEX, JOE and SIMA dyes due to more efficient fluorescence quenching by BHQ-1. We successfully used R6G dual-labeled probes for rotavirus genotyping.
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