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Agarose Gel Electrophoresis of DNA - Term Paper Example

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Agarose Gel Electrophoresis of DNA Introduction Electrophoresis refers to a method used to separate and purify macromolecules, mostly nucleic acids and proteins, which differ in conformation, size, or charge…
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Agarose Gel Electrophoresis of DNA
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Agarose Gel Electrophoresis of DNA Introduction Electrophoresis refers to a method used to separate and purify macromolecules, mostly nucleic acids and proteins, which differ in conformation, size, or charge. Gel electrophoresis is applied in clinical chemistry to separate proteins by size and/or charge and in molecular biology and biochemistry to separate RNA and DNA fragments by length, separating proteins by charge, or estimating the size of RNA and DNA fragments. To separate nucleic acid molecules, an electric field is applied to move molecules with negative charges through an agarose matrix. The short molecules move fast hence migrating farther compared to the longer ones. This is due to the molecule’s ability to easily migrate via the pores of the gel. A charge separates proteins in agarose due to the pores of the gel being too large to sieve them (proteins). Gel electrophoresis is also used in the separation of nanoparticles (Colorado State University, 1). Agarose Gel Electrophoresis of DNA in Context Agarose is a polysaccharide which is extracted from seaweed. Typically it is used at 0.5% to 2% concentrations. A high agarose concentration makes the gel stiffer, and vice versa. Agarose gels are easy to prepare and non toxic. Agarose powder is simply mixed with a buffer solution then melted by heating, and then the gel is poured. Agarose gels have a low resolving power, but relatively a large separation range. DNA fragments from around 200 to 50,000 base pair can be separated by varying agarose concentration using electrophoretic techniques. Read More
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