Gel Electrophoresis is a technique used to separate and identify

specific compounds in a mixture using an electrical field. In this procedure,

the technique will be used to separate fragments of deoxyribonucleic acid
(DNA). The negatively charged DNA molecule is pulled through the gel matrix
toward the positive pole. The purpose of this procedure is to separate DNA
strands for further analysis.
Our group received an agarose gel tray. We placed with a pipette two
dyes, 2 DNA markers, and an unknown DNA sample into the tray. This was
placed on the platform of the electrophoresis chamber. The casting chamber
was placed near the black electrode. Then approximately 2.5 liters of buffer
solution was added to the chamber. The lid was placed on the machine and it
was turned on. The buffer solution showed signed of bubbling. When it was
determined that the electrophoresis separation had occurred the power was
turned off. The tray was removed from the chamber. The agarose gel was
then stained in methylene blue to make the separated fragments visible.
This made three bands visible, blue, orange, and purple. The separation of
these three dyes shows that the lighter the molecule, the farther it will travel
through the agarose gel. The electrophoretic migration of the known,
standard DNA fragment is compared with an unknown fragment to identify
the length of the unknown fragment. This is done by measuring the distance
from the middle of the starting well to the leading edge of each band.
Each of our known DNA fragments has a known length and is
expressed in kilobase pair (KBP) value. The distance migrated in expressed in
millimeters. Our gel electrophoresis findings are; DNA marker one migrated
1.1 millimeters. DNA marker two migrated 1.3 millimeters. DNA marker three
migrated 1.5 millimeters. DNA Marker four migrated 1.9 millimeters. DNA
marker five migrated 2.6 millimeters. DNA maker six migrated 2.7
millimeters. The unknown DNA marker migrated 2.6 millimeters. Therefore
rendering our unknown to have a result of 2.6 KBP.
DNA Marker
Fragment
Number
1
2
3
4
5

DNA Marker
Fragment
Length (KBP)
23.13
9.41
6.68
4.36
2.32

DNA Marker
Fragment
Distance
Migrated (mm)
1.1
1.3
1.5
1.9
2.6

6
Unknown

2.03
2.5

2.7
2.6

These results have been plotted on a semi-log graph. The graph is

located on the following page.
Gel Electrophoresis is a process which enables the sorting of molecules
based on size. Using an electric field, DNA molecules can be made to move
through a gel made of agar or polyacrylamide. The electric field consists of a
negative charge at one end which pushes the molecules through the gel, and
a positive charge at the other end that pulls the molecules through the gel.
The molecules being sorted are dispensed into a well in the gel material. The
gel is placed in an electrophoresis chamber, which is then connected to a
power source. When the electric current is applied, the larger molecules
move more slowly through the gel while the smaller molecules move faster.
The different sized molecules form distinct bands on the gel. The standard
DNA fragments are identifiable and known markers. This is then compared to
the unknown DNA to determine the length in KBP. Gel Electrophoresis is
used to separate fragments of DNA, RNA, and proteins for further analysis.
Our unknown DNA fragment migrated 2.5 millimeters and the
fragments length was 2.6 KBP.