Gel electrophoresis is a technique used to separate macromolecules like proteins and nucleic acids based on size, charge, or conformation. It works by applying an electric field to push molecules through a gel containing small pores. Smaller molecules will travel farther through the gel than larger ones. After electrophoresis, the gel can be stained to make the separated molecules visible, then used for various applications like DNA fingerprinting in forensics, mapping cellular components in biochemistry, and determining purity in protein analysis.

1. Gel Electrophoresis
By
Harshita Jain
Assistant professor
Career Point School of pharmacy Career Point University, Kota

2. CONTENTS
Introduction
Principle Of Electrophoresis
Electrophoresis Apparatus
Procedure For Electrophoresis
Applications Of Electrophoresis

3. INTRODUCTION
Electrophoresis is a technique used to separate and some
time purify macromolecules – especially proteins and
nucleic acids- that differ in size , charge or conformation.
The migration of electrically charged particles or ions in
solutions due to an applied electric field.
In practical terms a positive electrode (anode) and negative
electrode(cathode) are placed in a solution containing ions.
Then voltage is applied across the electrodes, so solute ions
of different charge for example, anions (negative) and
cations (positive) will move through the solution towards
the electrode of opposite charge.

6. PRINCIPLE OF GEL
ELECTROPHORESIS
 Gel electrophoresis involves an electric field.
 The molecules to separated are pushed by an electrical
field through gel that contains small pores
 The molecules travel through the pores in the gel at
a speed that is inversely related to their lengths.
 This means that a small DNA molecule will travel a
greater distance through the gel than will a larger DNA
molecule
Electromigration :
• – Ions migrating in electric field
Cations cathode (-ve)
Anions  anode (+ve)

7. ELECTROPHORESIS
APPARATUS

9. VISUALISATION
• After the electrophoresis is complete, the molecules in the
gel can be stained to make them visible.
• Ethidium bromide, silver, or coomassie blue dye may be
used for this process.
• If the analyte molecules fluoresce under ultraviolet light, a
photograph can be taken of the gel under ultraviolet
lighting conditions.
• If the molecules to be separated contain radioactivity added
for visibility, an autoradiogram can be recorded of the gel.

10. APPLICATIONS
1. Forensics
• DNA fingerprint of acriminal.
2. Molecular Biology To separate and organize DNA and RNA
by size
3. Genetics Provide clearer picture of DNA, it also helps
prepare DNA for cloning and geneticengineering.
4. Microbiology Information out about the organisms.
Virology : to help diagnose different strains of viruses.
5. Biochemistry Mapping of cellular components, particularly
proteins and nucleic acids.

11. 6. Deter,minationofproteinandpeptide
(1) Tocheck Purity
(2) Physical properties like MW,pI
(3) Binding studies
(4) Identification (CE-MS)
(5) Quantitation
(6) Immunoassays
7. Determination of additives in food anddrug.
For example, Separation of Tartazine, Sunset yellow, amaranth,
indigo carmine
8.Separation and Quantification of Vitamins in fruits and
vegetable.
For example, Separation and quantification of Ascorbic acidin
vegetable and fruits