Solid- and solution-phase synthesis and application of R6G dual-labeled oligonucleotide probes.
Bioorg Med Chem
; 23(20): 6749-56, 2015 Oct 15.
Article
em En
| MEDLINE
| ID: mdl-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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1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Rodaminas
/
Sondas de Oligonucleotídeos
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article