ResourceSpectroscopy
Determination of Nucleic Acid Concentrations Using Fluorescent Dyes in the Eppendorf BioSpectrometer® fluorescence
14 Oct 2014Fluorescence allows more sensitive and more specific determination of nucleic acids than the conventional method of UV-VIS spectroscopy. The present article describes quantification of dsDNA using the fluorescent dye PicoGreen®, with special focus on the preparation of samples for measurement in the Eppendorf BioSpectrometer, as well as the details of programming. The methods PicoGreen and PicoGreen-“short” are compared experimentally. Even though the method PicoGreen-“short” was measured using only 2 standards, high analytical accuracy was observed.
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Fluorescence SpectroscopyFluorometers and spectrofluorometers (also called fluorescence spectrometers) are used to measure the intensity and wavelength of fluorescent light emitted from a sample after excitation by illumination. Spectrofluorometers utilize monochromators to select the desired wavelengths, whereas filter fluorometers employ a set of filters. Spectrofluorometers for measuring steady-state fluorescence and lifetime fluorescence (or time-resolved fluorescence) are available, as well as fluorescence microscopes and microplate readers. Find the best fluorescence spectroscopy products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA / RNA Extraction and PurificationPurified DNA and RNA are required for numerous downstream molecular biology applications. Consequently, the importance of high-quality DNA/RNA extraction and purification equipment cannot be underestimated. Many purification kits are available and are typically optimized for nucleic acid type and source, including plasmid DNA, genomic DNA, mRNA, RNA and viral nucleic acid purification kits. Automated extraction and purification of nucleic acids can be implemented with magnetic bead separator instruments or high-throughput purification workstations. Find the best DNA/RNA extraction and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.FluorescenceThe emission of fluorescence occurs when a photon of energy is supplied to a fluorescent chemical compound by an external source, causing it to become excited. Fluorescence can be detected and measured for different purposes using microplate readers, fluorescence microscopes, fluorescence scanners, and flow cytometers.Nucleic Acid Concentration