- DNA electrophoresis
DNA electrophoresis is an analytical technique used to separate
DNA fragments by size. Anelectric field forces the fragments to migrate through agel . DNA molecules normally migrate from negative to positive potential due to the net negative charge of thephosphate backbone of the DNA chain. At the scale of the length of DNA molecules, the gel looks much like a random, intricate network. Longer molecules migrate more slowly because they are more easily 'trapped' in the network.After the separation is completed, the fractions of DNA fragments of different length are often visualizing a
fluorescent dye specific for DNA, such asethidium bromide . The gel shows bands corresponding to different DNA molecules populations with different molecular weight. Fragment size is usually reported in "nucleotides", "base pairs" or "kb" (for 1000's of base pairs) depending upon whether single- or double-stranded DNA has been separated. Fragment size determination is typically done by comparison to commercially availableDNA ladder s containing linear DNA fragments of known length.The types of gel most commonly used for DNA electrophoresis are agarose (for relatively long DNA molecules) and polyacrylamide (for high resolution of short DNA molecules, for example in
DNA sequencing ). Gels have conventionally been run in a "slab" format such as that shown in the figure, butcapillary electrophoresis has become important for applications such as high-throughput DNA sequencing. Electrophoresis techniques used in the assessment ofDNA damage includealkaline gel electrophoresis andpulsed field gel electrophoresis . The measurement and analysis are mostly done with a specialized [http://www.labimage.net gel analysis software] . Capillary electrophoresis results are typically displayed in a trace view called anelectropherogram .The DNA strand is cut into smaller fragments using a DNA restriction endonuclease, then samples of the DNA solution (DNA sample and buffer) are placed in the wells of the gel and allowed to run for some time (the less the voltage of the electophoresis, the longer time for the DNA sample to run through the gel, and this results in a more accurate separation).
External links
* [http://www.gel-electrophoresis.com/gels/articles/dna-gel-electrophoresis/ DNA Gel Electrophoresis]
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