Tip:
Highlight text to annotate it
X
Agarose Gel Electrophoresis Agarose gel electrophoresis is the most commonly
used method in the field of biochemistry and molecular biology for the separation of DNA.
This technique also supports the separation and analysis of proteins and RNA.
Agarose is a polysaccharide from the red algae, is used for a variety of life science applications
especially in gel electrophoresis. Agarose forms an inert matrix utilized in separation
techniques. Many structures easily affix to agarose including various types of proteins.
Electrophoresis is a technique used to separate and sometimes purify macromolecules - that
differ in size, charge or conformation. The equipment and supplies necessary for conducting
agarose gel electrophoresis are relatively simple.
An electrophoresis chamber and power supply Gel casting trays, which are available in
a variety of sizes and composed of UV-transparent plastic..
Sample combs, around which molten agarose is poured to form sample wells in the gel.
Electrophoresis buffer, usually Tris-acetate-EDTA (TAE) or Tris-borate-EDTA (TBE).
Loading buffer, which contains something dense (e.g. glycerol) to allow the sample to "fall"
into the sample wells, and one or two tracking dyes, which migrate in the gel and allow visual
monitoring or to indicate how far the electrophoresis has proceeded.
Ethidium bromide, a fluorescent dye used for staining nucleic acids and it helps to visualize
the DNA molecule.The property of this chemical is that it binds to the DNA bases strongly
and flouresces orange. Separat ion of the molecules is achieved by
the movement of negatively charged nucleic acid molecules through an Agarose matrix in
an uniform electric field . The movement depends on the length and conformation of the molecule.
Movement of DNA.
Transilluminator (an ultraviolet light box), which is used to visualize ethidium bromide-stained
DNA in gels.
The main benefit of Agarose gel electrophoresis is that it can be a preparative technique
as the DNA can also be recovered without any harm to it at the end of the process. Agarose
gel does not denature the DNA samples and stay in their native form.
Gel Electrophoresis can be used in the field of forensics. The process of DNA finger printing
can be performed using the Agarose gel DNA electrophoresis.