Answer 1:
Great questions! Electrophoresis is a technique
used by many scientists, but not very well known
by many students, so thanks for asking! Gel
Electrophoresis is basically a technique used to
separate nucleic acids (the material of DNA and
RNA) or proteins according to their relative
sizes. Say a scientist has a tube with a mixture
of different sizes of DNA or protein, and would
like to separate one size or one piece of this
mixture away from the rest of the sizes or pieces.
The best way to do this is gel
electrophoresis. The term electrophoresis
comes from electro- and phoresis. "Phoresis" means
basically "movement" and "electro" refers to the
electrical field that's used to separate the
nucleic acids or proteins. In overview; samples
are run through a gel network, moved by an
electric field, and will separate according to
size. After the samples are separated, they can be
analyzed. The gel serves as a solid mesh to
support the biological sample as is it separated
by the electrical field. The gel is like a kitchen
sponge, but with a microscopic network of holes
and connections. For DNA samples, generally a gel
is made like you would make jell-o, by heating up
a polymer (agarose) with liquid, poring it into a
mold, and allowing the network to solidify. (Fun
fact: agarose is made from seaweed, and is
actually in some products like toothpaste and
gummy snacks!) For protein samples, generally a
finer network (smaller holes) is required, so a
gel is made of a different polymer (acrylamide).
Once the gel has solidified, it is covered
in a liquid (buffer) through which an electric
current can pass. Samples are set into the gel,
usually in small niches, or "wells". The protein
or nucleic acid will generally be mixed with a dye
to see the samples, and a component to help the
sample sink in the well, and to not mix with the
(buffer). An electric current is applied to the
gel and samples, by having a current applied to a
cathode nearest to the samples, and an anode at
the other end of the gel. Nucleic acids are
negatively charged, and will move to the
positively charged end of the gel. Proteins have
different shapes and charges, and are treated
before electrophoresis to have approximately the
same shape (straightened out) and uniform negative
charge. As the sample moves through the
gel, the mix of sizes starts to be separated. You
can imagine a group of creatures running through a
dense forest; the smallest critters can maneuver
the fastest through the trees, while the giants
take much longer to navigate through! After
the sample has run the length of the gel, the
electrical force is stopped, and the samples are
visualized as "bands" or short lines on the gel,
kind of like a ladder. A sample of known sizes in
run simultaneously with the other samples. This
allows for comparison and determining the
approximate size the unknown sample bands. Nuclei
acids can be visualized by staining with a
molecule (ethidium bromide) that slots between the
nucleic acid "base pairs" and will glow under
ultra violet light (like a black light). Protein
is generally stained with a dye. The separated
sample bands can then be photographed, transferred
to another material to be further characterized,
or single bands can be cut out from the gel and
the sample purified from the gel! Gel
Electrophoresis is used in many fields, including
molecular biology, biochemistry, forensics, and
genetics; it is used frequently in health
diagnosis. The compounds used in this
technique can be toxic, and the electrical field
could be harmful if not used properly. But, if
people are trained, and have proper supervision
and equipment, they could run a successful gel
electrophoresis. In certain programs and classes,
high school students frequently learn and apply
this technique! Click Here to return to the search form.
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