Gel electrophoresis is a method used to separate molecules like proteins and nucleic acids based on their physical properties. It works by applying an electric field to move charged molecules through a gel medium. Key aspects include applying samples to the gel, running electrophoresis with an electric current, and visualizing separated molecules by staining and quantification using a densitometer. Gel electrophoresis has applications in forensics, genetics, and biochemistry to analyze DNA, diagnose diseases, and study proteins. It separates biomolecules based on factors like size, shape, and charge.

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

3. ELECTROPHORESIS
 Electrophoresis is the migration of charged particles 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
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4. PRINCIPLE
 Electrophoresis is the process of migration of charged
molecules through solution in an applied electric field.
 Electrophoresis is often classified according to the
presence or absence of a solid supporting medium or
matrix through which the charged molecules moves in
the electrophoretic system.
 Solution electrophoresis system employs aqueous
buffers in the absence of solid support medium. Such
system can suffer from sample mixing due to diffusion
of the charged molecules with resolution loss of
resolution during the sample application separation &
removal steps.
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5. Separation of molecules on the
basis of
>size and/or
>charge and/or
>shape
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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 ,
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8. APPLICATIONS
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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.
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11.  APPICATION OF SAMPLE
 RUNNING OF SAMPLE
 VISUALISATION OF SAMPLE
 QUANTIFICATION
TECHNIQUES
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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.
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13. DENSO METER
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14. 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. 14

15. RANNING ELECTROPHORESIS
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16. Gel electrophoresis is a
process that separates
fragments of DNA based on
their sizes.
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17. 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
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18.  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
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19. 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
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20. REFERENCE
 colloidal chemistry by G Whitmore published by
prabhath Kumar Sharma or sarup and sons page
no:44
 colloidal chemistry by B K Sharma page no:84
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