This document provides an introduction to gel electrophoresis. Gel electrophoresis is a method used to separate molecules like proteins and nucleic acids based on physical characteristics like size, shape, and charge. It works by applying an electric field to move molecules through a gel matrix. There are different types of gels that can be used like agarose or polyacrylamide. The document discusses the basic procedure, which involves applying samples to a gel, running an electric current, staining to visualize the separated molecules, and analyzing the results. Applications of gel electrophoresis include determining DNA fragment sizes, solving criminal cases or determining paternity or species relationships.

1. WELCOME

2. GEL ELECTROPHORESIS
Guided by:
Ms. Ashwini D
Dept. of Industrial
chemistry
Presented by:
Suresh B I

3. ELECTROPHORESIS
 Electrophoresis is the migration of charged particle in an electric
field towards the electrode bearing the opposite charge.
(OR)
 is a method where by charged molecules in solution, like
proteins and nucleic acids migrate in response to an
electric field

4. PRINCIPLE
powerful separation method frequently used to
analyze DNA fragments generated by restriction
enzymes
convenient analytical method for determining the
size of DNA molecules in the range of 500 to 30,000
base pairs.
employs electromotive force to move molecules
through a porous gel

5.  separates molecules from each other on
the basis of
>size and/or
>charge and/or
>shape
 basis of separation depends on how the
sample and gel are prepared

7. TYPES OF
ELECTROPHORESIS
Based on supporting
media (zone
electrophoresis):
 Paper electrophoresis
 gel electrophoresis
 Cellulose acetate
electrophoresis
Without supporting
media (moving boundary
method):
 Free electrophoresis
Supporting medias are :
starch, Agar- Agarose,
polyacrylamide ,

8. APPLICATIONS

10. GEL ELECTROPHORESIS
Introduction: Gel electrophoresis is group of
techniques used by scientists to separate molecules
based on physical characteristics such as size, shape,
and isoelectric point. Gel electrophoresis is usually
performed for analytical purposes.

11.  APPICATION OF SAMPLE
 RUNNING OF SAMPLE
 VISUALISATION OF SAMPLE
 QUANTIFICATION
TECHNIQUES

12. REQUIREMENTS FOR GEL
ELECTROPHORESIS
 Buffer
 Fixative
 Staining solution
 Destaining solution
 Densitometer – is essentially a double beam filter
photometer or spectrophotometer that scans the
electrophoretic strip ( in the form of agarose , cellulose
acetate or polyacrylamide) as it moves past the optical
system.

13. What is Agarose ?
A linear carbohydrate polymer extracted from seaweed
, agarobiose
forms a porous matrix as it gels
shifts from random coil in solution to structure in which
chains are bundled into double helices

14. DENSO METER

15. PROCEDURE
1. Sample to be separated is applied to a supporting
medium (paper, cellulose acetate, agar gel, polyacrylamide gel etc.)
Electrophoresis is carried out at desired constant voltage or
constant current in presence of specific pH.
3. After completion of electrophoresis the supporting
medium is placed in a fixative to prevent diffusion of
separated fractions.
4. Separated fraction is then visualized by using appropriate stains
e.g. Bromophenol Blue & Amino Schwartz for plasma proteins and
Sudan Black for lipoproteins.
5. Quantification of each fraction is done by either densitometer
or elution followed by colorimeter or spectrophotometer of
eluted fraction.

16. RANNING ELECTROPHORESIS

17. Gel electrophoresis is a
process that separates
fragments of DNA based on
their sizes.

18. APPLICATION OF GEL
ELECTROPHORESIS
 Evidence in criminal cases
 To determine paternity
 To diagnose genetic diseases
 Help to determine kinship in animals
 Compare similarities and differences between
species
Solve paternity cases
Determine genetic kinship among species

19.  Estimated molecular masses and relative abundance of
unknown polypeptides in a complex mixture
 Patterns of bands that suggest presence of isoenzymes or
specific complex proteins
 Effectiveness of a separation procedure during cell/tissue
fractionation
 Effectiveness of a procedure to purify specific organelles,
proteins, or polypeptides

20. CONCLUSION
 Gel electrophoresis is used in forensic, molecular
biology and bio chemistry
 The results can be analysed quantitatively by
visualizing the gel with Uv light and a gel imaging
device
 The image is recorded with a computer operated
camera and the intensity of the band (or) spot of
interest is measuring and compared against standard
(or)markers loaded on the sample gel
 The measurement and analysis are mostly done
with specialised software's

21. REFERENCE
 colloidal chemistry by G Whitmore published by
prabhath Kumar Sharma or sarup and sons
 colloidal chemistry by B K Sharma

22. THANK YOU