Introduction to Electrophoresis & types

Electrophoresis is extensively utilized in a range of sectors, such as genetics, biochemistry, and forensic science, for purposes such as DNA analysis, protein purification, and investigating gene expression.
Its crucial role in modern biological research lies in its capability to separate and analyze complex mixtures effectively.

1. Rinde Usama Kasam

2. Introduction to Electrophoresis & its types
Electrophoresis.
Factors influencing ion
migration in electric
field.
Types of electrophoresis.
Paper Electrophoresis.
Gel Electrophoresis.
Capillary Electrophoresis.
References

3. Introduction to Electrophoresis
Electrophoresis involves the migration of
charged particles or molecules in a medium
due to an electric field.
It is a broad term that describes the
movement of charged particles in a liquid
medium when exposed to an electric field.
The direction in which molecules move
depends on their charge, whether they are
positive or negative.
An ampholyte becomes positively charged in
acidic conditions and moves towards the
cathode, while in alkaline conditions they
become negatively charged and move towards
the anode
It is a more cost-effective separation method
as opposed to the alternatives.
Electrophoresis is commonly utilized to
analyze RNA, DNA, and proteins.
It is frequently employed for the isolation of
amino acids and proteins.

4. Factors influencing ions migration in an electric field

Electric charge of molecule
Particle dimensions and form
Intensity of electric field
Characteristics of the surrounding environment
Operating temperature range

5. Different forms of Electrophoresis

Free Electrophoresis: It is a technique that is carried out without
the use of supporting media.
Zone Electrophoresis: Electrophoresis in zone is done by
utilizing support materials such as paper or gel for separation. The
parts are divided into zones, which is why they are called that way
& It is also referred to as electromatography.
Gel-Electrophoresis: The Agarose gels and sodium dodecyl
polyacrylamide are commonly employed as support matrices in this
application. The method is best suited for isolating proteins & It
can serve for analytical purposes and as a preparative method for
purifying molecules beforehand applying different analytical
methods.
Capillary Electrophoresis: It involves the use of small diameter
tubes to separate the samples. It is a straightforward, fast, simple,
and more effective approach that uses a smaller sample.

6. Paper Electrophoresis
This method may come in handy for
isolating amino acids, small proteins, or
small molecules with a charge.
The paper strips are wet with buffer
solution and one end of the strip is
immersed in buffer solution that
contains an electrode.
Apply the sample at the middle of the
paper and apply high voltage.
The specimen will move based on their
charges.
After conducting electrophoresis, the
isolated components can be recognized
using different staining methods based
on their chemical composition.

7. Gel Electrophoresis

Similar to the other electrophoresis methods,
this technique also utilizes an electric field.
The Phytoconstituents that need to be
separated are added to the gel while an
electric field is applied.
The gel includes the pore that
Phytoconstituents must pass through.
Phytoconstituents small molecules will
travel a greater distance due to their ability
to penetrate tiny pores in the gel allow
smaller molecules to pass through more
easily than larger molecules.
In general, DNA and RNA, both carrying a
negative charge, can be effectively separated
through gel electrophoresis.
To separate proteins using this method, they
are initially exposed to sodium dodecyl
sulfate which straightens out the protein and
covers it with a negative charge.
Agarose gel or polyacrylamide gel are
currently utilized for gel electrophoresis.

8. Capillary Electrophoresis

During capillary electrophoresis, separation occurs within
a capillary with an internal diameter ranging from 10 to
100 µm.
The tips of the fused silica capillary tube are immersed in
the buffer solution.
It includes platinum electrodes (both cathode and anode).
Capillary tube has an optical window for detection that is
connected to a UV detector.
In order to introduce the sample, the inlet buffer must be
swapped for the sample, which can be injected using
pressure, electro-kinetically, capillary action, or siphoning.
When voltage is applied, the sample begins to move, a
process called electrophoretic migration, which is also
influenced by the buffer solution's electron osmotic flow.
Typically, the electro osmotic movement is directed
towards the cathode (or negatively charged).
While electro osmotic flow has a higher electrophoretic
mobility, all the analytes are still transported towards the
cathode at various speeds.
Detector can detect these.

9. References
Dr.K Prabhu, D. (2019). Pharmacognosy & Phytochemistry 2.
UP: Thakur Publication.
Dr.Parbodh Shukla, D. S. (2019). Pharmacognosy &
Phytochemistry 2. UP: Nirali Prakashan.