Shree Kushwaha

Date: 02/26/2013

Gel-Electrophoresis

Claflin University
Biology Department
Dr. Ryan Kendal
Genetics Lab Instructor
 Abstract
The main objective of the today’s experiment was to learn the gel electrophoresis
technique to separate the different strand of the DNA molecules cut by the
respective restriction enzyme. Since DNA is negatively charged molecules, in an
electric current it moves from a negative pole to a positive pole. From the gel
pictures it was found that the smaller strand of DNA molecules moves faster than
larger strand. The size of the DNA strand was measured on the basis of base pairs
present in that specific strand.
 Introduction
Gel electrophoresis is a biological technique used to separate fragments of DNA
according to size. In order to obtain the fragments of DNA, DNA is treated with
the restriction enzymes. Upon electrical stimulation, smaller fragments of a
molecule will move faster through the gel than larger fragments. Since DNA is
negatively charged molecules it moves from the negative pole of the electricity to
the positive pole. Ladder is used to measure the size of the DNA fragments.

Using a pipette, DNA samples are loaded into wells made in the agarose
gel. Since DNA samples are colorless researchers add blue “tracking” dye. This
makes it easier to load the samples, and visually track the DNA migration through
the gel. The phosphate groups in the DNA backbone carry negatively charged
oxygen- giving a DNA molecule negative charge. In an electric current, the
negatively charged DNA moves toward the positive pole of the electrophoresis
chamber. After the gel has run for certain period of time, gel is stained with
ethidium bromide. Ethidium bromide binds tightly to the DNA, and glows when
illuminated with UV light. This lets researchers see where the separate DNA
fragments end up.

In today’s lab our DNA samples was cut with three different restriction enzymes
namely, BamHI, EcoRI, and HindIII. These three samples were loa
 Materials and Methods
Gel, digested DNA, ladder was provided by the instructor. The samples were
loaded in the wells along with marker and a control. After loading the samples the
gel was run for about 40-50 minutes by passing the current. After 50 minutes the
electricity was disconnected, and a gel picture were taken. The size of the
fragments was analyzed by comparing with size of the marker.
 Results

DN
A
Lad

10
8
6
5
4
3
2.5
2
1.5

From the above gel electrophoresis pictures we can analyzed that we have run five
samples. The first one is marker also called as Ladder, second one DNA samples
cut with Bam HI restriction enzymes, the third one is cut with Eco RI, fourth one is
cut with Hind III, and last one is just DNA samples without any restriction enzyme
working as control. So basically we can say that all of the samples above are single
digested, cut with the single restriction enzymes. The size of fragments for each
sample is compared with the size of the marker.
 Conclusion and Discussion
In today’s experiment the basic the concept and technique of gel electrophoresis
was learned. The restriction enzyme was used to cut the DNA molecules into
different pieces. DNA samples were loaded in the gel and were run on the gel. It
was found that smaller strands of DNA molecules moves faster than the larger
strand. In this way different strand of DNA molecules were separated depending
on the size. The size of the DNA strand is measured on the basis of the base pairs
present in the specific strand.
 References
1. http://www.life.illinois.edu/molbio/geldigest/electro.html. Web.
28 Mar. 2013.

2. Bowen. "Agarose Gel Electrophoresis of DNA." Agarose Gel

Electrophoresis of DNA. N.p., 2000. Web. 28 Mar. 2013.

3. http://www.sumanasinc.com/webcontent/animations/content/ge
lelectrophoresis.html. Web. 28 Mar. 2013.