Solid- and solution-phase synthesis and application of R6G dual-labeled oligonucleotide probes

Aleksander Yu Skoblov, Maxim V. Vichuzhanin, Valentina M. Farzan, Olga A. Veselova, Tatiana A. Konovalova, Alexander T. Podkolzin, German A. Shipulin, Timofei S. Zatsepin

    Research output: Contribution to journalArticlepeer-review

    7 Citations (Scopus)

    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.

    Original languageEnglish
    Pages (from-to)6749-6756
    Number of pages8
    JournalBioorganic and Medicinal Chemistry
    Volume23
    Issue number20
    DOIs
    Publication statusPublished - 15 Oct 2015

    Keywords

    • CuAAC
    • Fluorescent dye
    • Oligonucleotide
    • Phosphoramidite
    • qPCR
    • Rhodamine 6G

    Fingerprint

    Dive into the research topics of 'Solid- and solution-phase synthesis and application of R6G dual-labeled oligonucleotide probes'. Together they form a unique fingerprint.

    Cite this