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Gel electrophoresis of nucleic acids is an analytical technique to separate DNA or RNA fragments by size and reactivity. Nucleic acid molecules are placed on a gel, where an electric field induces the nucleic acids (which are negatively charged due to their sugar-phosphate backbone) to migrate toward the positively charged anode. The molecules ...
Gel electrophoresis of large DNA or RNA is usually done by agarose gel electrophoresis. See the "chain termination method" page for an example of a polyacrylamide DNA sequencing gel. Characterization through ligand interaction of nucleic acids or fragments may be performed by mobility shift affinity electrophoresis.
A number of factors can affect the migration of nucleic acids: the dimension of the gel pores (gel concentration), size of DNA being electrophoresed, the voltage used, the ionic strength of the buffer, and the concentration of intercalating dye such as ethidium bromide if used during electrophoresis.
Nucleic acids or nucleic acid fragments may be characterized by their affinity to other molecules. The methods have been used for estimation of binding constants , as for instance in lectin affinity electrophoresis or characterization of molecules with specific features like glycan content or ligand binding. [ 1 ]
Polyacrylamide gel electrophoresis (PAGE) is a technique widely used in biochemistry, forensic chemistry, genetics, molecular biology and biotechnology to separate biological macromolecules, usually proteins or nucleic acids, according to their electrophoretic mobility. Electrophoretic mobility is a function of the length, conformation, and ...
Gel electrophoresis: The DNA fragments are then electrophoresed on an agarose gel to separate them by size. If some of the DNA fragments are larger than 15 kb , then before blotting, the gel may be treated with an acid, such as dilute HCl .
Agarose gel electrophoresis is the routine method for resolving DNA in the laboratory. Agarose gels have lower resolving power for DNA than acrylamide gels, but they have greater range of separation, and are therefore usually used for DNA fragments with lengths of 50–20,000 bp ( base pairs ), although resolution of over 6 Mb is possible with ...
GelGreen is an intercalating nucleic acid stain used in molecular genetics for agarose gel DNA electrophoresis. GelGreen consists of two acridine orange subunits that are bridged by a linear oxygenated spacer. [1] [2] Its fluorophore, and therefore its optical properties, are essentially identical to those of other N-alkylacridinium orange dyes.