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Agarose
gel electroporesis
Agarose
gel electrophoresis separates DNA fragments according to their size. Typically,
a DNA molecule is digested with restriction
enzymes,
and the agarose gel electrophoresis is used as a diagnostic tool to visualize
the fragments. An electric current is used to move the DNA molecules across
an agarose gel, which is a polysaccharide matrix that functions as a sort
of sieve to help "catch" the molecules as they are transported by the
electric current. This
technique has lots of applications. Generally speaking you can analyze
DNA fragments that result from an enzyme digestion of a larger piece of
DNA to visualize the fragments and determine the sizes of the fragments.
In addition to its usefulness in research techniques, agarose gel electrophoresis
is a common forensic technique and is used in DNA fingerprinting.
The phosphate molecules that make up the backbone of DNA molecules have a high negative charge. When DNA is placed on a field with an electric current, these negatively charged DNA molecules migrate toward the positive end of the field, which in this case is an agarose gel immersed in a buffer bath. The agarose gel is a cross-linked matrix that is somewhat like a three-dimensional mesh or screen. The DNA molecules are pulled to the positive end by the current, but they encounter resistance from this agarose mesh. The smaller molecules are able to navigate the mesh faster than the larger one, so they make it further down the gel than the larger molecules. This is how agarose electrophoresis separates different DNA molecules according to their size. The gel is stained with ethidium bromide so you can visualize how these DNA molecules resolved into bands along the gel. Southern blotting may also be used as a visualization technique for agarose gels.
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