A Solution to the Common Problem of the Synthesis and Applications of Hexachlorofluorescein Labeled Oligonucleotides
| Autor: | Galina E. Pozmogova, Alena G. Mosina, Anna M. Varizhuk, Igor P. Smirnov, A. N. Chuvilin |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Reversed-Phase High Performance Liquid Chromatography Deblocking filter Oligonucleotides lcsh:Medicine Artificial Gene Amplification and Extension Biochemistry Polymerase Chain Reaction Physical Chemistry chemistry.chemical_compound Heterocyclic Compounds Solvolysis lcsh:Science Liquid Chromatography Multidisciplinary Chemistry Organic Compounds Nucleotides Chromatographic Techniques Chemical Reactions Photochemical Processes Physical Sciences Research Article Fluorophore Photochemistry Side reaction Research and Analysis Methods 03 medical and health sciences Ammonia Molecular Biology Techniques Molecular Biology Fluorescent Dyes 030102 biochemistry & molecular biology Staining and Labeling Oligonucleotide Spectrum Analysis lcsh:R Organic Chemistry Chemical Compounds Biology and Life Sciences Reversed Phase Chromatography Combinatorial chemistry High Performance Liquid Chromatography Electrophoresis 030104 developmental biology Yield (chemistry) Acridines lcsh:Q Derivative (chemistry) |
| Zdroj: | PLoS ONE PLoS ONE, Vol 11, Iss 11, p e0166911 (2016) |
| ISSN: | 1932-6203 |
| Popis: | A common problem of the preparation of hexachlorofluorescein labeled oligonucleotides is the transformation of the fluorophore to an arylacridine derivative under standard ammonolysis conditions. We show here that the arylacridine byproduct with distinct optical characteristics cannot be efficiently separated from the major product by HPLC or electrophoretic methods, which hampers precise physicochemical experiments with the labeled oligonucleotides. Studies of the transformation mechanism allowed us to select optimal conditions for avoiding the side reaction. The novel method for the post-synthetic deblocking of hexachlorofluorescein-labeled oligodeoxyribonucleotides described in this paper prevents the formation of the arylacridine derivative, enhances the yield of target oligomers, and allows them to be proper real-time PCR probes. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|