TAHSIN HOSSAIN SAMIK SAHA HUMAYRA AFRIN

ID: 20052055 ID: 20052065 MIM

ID: 20052076
 PRESENTATION ON

AGAROSE GEL
ELECTROPHORESIS
WHY WE DO IT?

Agarose Gel Electrophoresis separating

DNA fragments based on their molecular
size, which is crucial in genetic research
and medical diagnostics.
What We Need for Agarose Gel Electrophoresis?

Forms the gel

matrix for molecule The specimens
separation to be separated
and analyzed

Agarose powder DNA Samples

Enhances sample
visibility and
Conducts current
facilitates loading
and maintains pH into wells

Electrophoresis Buffer Loading Dye

What We Need for Agarose Gel Electrophoresis?

Holds the gel

A marker for during
estimating DNA preparation and
fragment sizes electrophoresis

DNA Ladder Gel Casting Tray

Comb

Provides the Forms wells in

Visualizes DNA the gel for DNA
electric current
bands after sample loading
supply for DNA
electrophoresis
migration
Power Supply UV Transilluminator
 PROCEDURE OF AGAROSE GEL ELECTROPHORESIS

Creating the Gel: Agarose and Buffer

1. Dilute concentrated 50X Electrophoresis buffer
Agarose
with distilled water. Weigh 2 grams of agarose Concentrated
Buffer
Distilled Water

powder and add it to 100 ml of buffer solution

(e.g., TAE or TBE) in a flask.

2. Heat the mixture until the agarose dissolves Heat

completely.

3. Cool agarose to 60°C with careful swirling. Pour

the liquid agarose solution into the gel casting tray.
Wait

POUR

Casting tray
 PROCEDURE OF AGAROSE GEL ELECTROPHORESIS

Creating Wells: The Starting Line for DNA

1. Insert the comb into the gel

to create wells.

2. Allow the gel to solidify at

room temperature.
 PROCEDURE OF AGAROSE GEL ELECTROPHORESIS

Micropipette
Loading DNA Samples: The Race Begins

1. Mix each DNA sample with an appropriate

volume of 1X loading buffer.

2. Carefully load the prepared samples into the

wells of the gel using a micropipette.

3. Include a DNA ladder in one of the wells for size

reference.
 PROCEDURE OF AGAROSE GEL ELECTROPHORESIS

Running the Gel: DNA on the Move

1. Connect the gel tank to the power supply.

2. Set the voltage to a constant value

(typically 80-120V) and run the gel for a
specified time according to the desired
separation.

3. After electrophoresis is complete, remove

the gel and casting tray from the
electrophoresis chamber.
 PROCEDURE OF AGAROSE GEL ELECTROPHORESIS

Visualizing Results: The Finish Line

1. Stain the gel with ethidium

bromide solution (final
concentration: 0.5 μg/ml) or other
dye for DNA visualization.

2. Visualize the DNA bands under

UV light using a transilluminator.
What do we see?

Smaller fragments migrate

faster than larger ones.