ResourceSpectroscopy
Comparison of Fluorescence-Based Quantitation with UV Absorbance Measurements
13 Aug 2015Detection and quantitation of nucleic acids are vital to many biological studies but the methods commonly used can be unreliable and inaccurate. UV absorbance measurements are not selective and cannot distinguish DNA, RNA, or protein and walues are easily affected by other contaminants and variations in base composition. In addition, the sensitivity of spectrophotometry is often inadequate, prohibiting quantitation of DNA and RNA at low concentrations. This application note compares the two quantitation methods using the Qubit® Fluorometer and a microvolume UV spectrophotometer. The Qubit® fluorometric quantitation provided a more selective, sensitive, and accurate method for quantitating nucleic acids than UV absorbance measurements.
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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.Light MicroscopyLight microscopes or optical microscopes are used to visualize microscale objects under magnification, including cells, clinical specimens and materials. Lab equipment for light microscopy includes confocal microscopes, fluorescence microscopes, zoom and stereo microscopes. Microscope slides and imaging reagents are available for visualizing samples, as well as various microscope stages and incubators for large or temperature-sensitive samples. Find the best light microscopes in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.DNA / RNA QuantificationDetection and quantification of nucleic acids is important in molecular biology, cloning, expression, forensics and clinical diagnostics. Nucleic acids can be detected by labeling with colorimetric, fluorescent or radio labels and using in situ hybridization kits to identify specific sequences. Multiple nucleic acids can be detected and quantified at once using RNA / DNA detection beads or RNA / DNA microarrays. Find the best DNA / RNA Quantification products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Nucleic Acid ConcentrationFluorescenceThe 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.DNA QuantificationDNA quantification is the process of measuring the amount of DNA in a sample. It is vital in genomics, molecular biology, and biotechnology, helping to ensure accurate experiments and data analysis. Explore DNA quantification tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.SpectrometrySpectrometry is a technique used to measure the mass-to-charge ratio of ions. It provides valuable information about the composition, structure, and molecular weight of compounds. Used across a variety of fields, including chemistry, biology, and environmental science, spectrometry is essential for identifying substances, analyzing mixtures, and developing new products. Explore the best spectrometry tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.UVUltraviolet (UV) light is used in a variety of applications, including disinfection, spectrophotometry, and molecular biology. UV light can damage DNA, making it a powerful tool in sterilization and in studies of mutagenesis and gene repair. Explore UV-related products in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.Photometry