ABSTRACT
We developed an efficient, cost-effective, and rapid purification method for chemically-labeled oligonucleotides that requires less time than conventional procedures such as ethanol precipitation or size-exclusion chromatography. Based on the hydrophilic and hydrophobic properties of DNA and amine-reactive fluorophores, we show that n-butanol saturated with distilled water may be used to remove unreacted fluorophores by sequestering them in the organic phase, while labeled DNA remains in the aqueous phase. This phase extraction method is simple, fast, and allows for processing multiple samples simultaneously, a necessity for high-throughput labeling strategies.
Subject(s)
Fluorescent Dyes/isolation & purification , Liquid-Liquid Extraction/methods , Oligonucleotides/isolation & purification , 1-Butanol/chemistry , Acetonitriles/chemistry , Carbocyanines/chemistry , Fluorescent Dyes/chemistry , Oligonucleotides/chemistry , Spectrometry, FluorescenceABSTRACT
UV irradiation of trans-resveratrol leads to its photochemical transformation to a new, highly fluorescent compound, whose chemical structure was unambiguously identified. The new compound has large values of fluorescence quantum yield, Stokes' shift, and two-photon absorption cross section, which make it suitable for bio-imaging, multi-color labeling, and two-photon microscopy.
