Introduction to Electrophoresis and its Classifications
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ELECTROPHORESIS-mobile.pptx
Electro refers to the energy of electricity and phoresis, means to carry across.
Electrophoresis is a technique for separating charged molecule in a mixture under the influence of an applied electric field.
PAPER ELECTROPHORESIS.pptx
Electrophoresis is a scientific laboratory technique that is used to separate DNA, RNA, or protein molecules based on their size and electrical charge. An electric current is passed through the molecules to move them so that they can be separated via a gel. The pores present in the gel work like a sieve, allowing smaller molecules to pass through more quickly and easily than the larger molecules. According to the way conditions are adjusted during electrophoresis, the molecules can be separated in the desired size range.
What is electrophoresis and what are its uses?
Electrophoresis is a very broadly used technique that, fundamentally, applies electric current to biological molecules – they’re usually DNA, but they can be protein or RNA, too – and separates these fragments into pieces that are larger or smaller in size.
The phenomenon of electrophoresis was first observed by Russian professors Peter Ivanovich Strakhov and Ferdinand Frederic Reuss in 1807 at Moscow University. A constant application of electric field caused the particles of clay dispersed in water to migrate, showing an electrokinetic phenomenon.
Electrophoresis can be defined as an electrokinetic process that separates charged particles in a fluid using an electrical field of charge. Electrophoresis of cations or positively charged ions is sometimes referred to as cataphoresis (or cataphoretic electrophoresis). In contrast, sometimes, the electrophoresis of anions or negatively charged ions is referred to as anaphoresis (or anaphoric electrophoresis).
It’s used in a variety of applications. Though it is most often used in life sciences to separate protein molecules or DNA, it can be achieved through several different techniques and methods depending upon the type and size of the molecules.
The methods differ in some ways, but all we need is a source for the electrical charge, a support medium and a buffer solution. Electrophoresis is also used in laboratories for the separation of molecules based on their size, density and purity.
The method used to separate macromolecules such as DNA, RNA, or protein molecules is known as gel electrophoresis.
It is used in forensics for –
Nucleic acid molecule sizing
DNA fragmentation for southern blotting
RNA fragmentation for northern blotting
Protein fragmentation for western blotting
Separation of PCR products analysis
Detection and analysis of variations or mutations in the sequence
Its clinical applications involve –
Serum protein electrophoresis
Lipoprotein analysis
Diagnosis of haemoglobinopathies and hemoglobin A1c.
The fundamental principle of electrophoresis is the existence of charge separation between the surface of a particle and the fluid immediately surrounding it. An applied electric field acts on the resulting charge density, causing the particle to migrate and the fluid around the particle to flow.
It is the process of separation or purification of protein molecules, DNA, or RNA that differ in charge, size.
Electrophoresis
This presentation provides an overview of electrophoresis. It begins by defining electrophoresis as the differential migration of ionized molecules in an electric field based on their charge-to-mass ratio. It then discusses the basic principles, factors affecting mobility, supporting media like paper and gels, common techniques including low voltage and capillary electrophoresis, and applications for analyzing proteins, DNA, antibiotics and more. The presentation concludes that electrophoresis is a useful method for separating charged substances, from small ions to large molecules, and is widely used despite some limitations.
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Electrophoresis is a separation Technique that is based on the movement of charged particles in an electric field.
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2. Active dielectrics readily store electrical energy, like piezoelectric and ferroelectric materials. Passive dielectrics restrict electrical flow and act as insulators, like glass and rubber.
3. When dielectric materials are placed in an electric field, dipoles are induced through mechanisms like electronic, ionic, orientational and space charge polarization, contributing to the material's dielectric properties. The different polarization mechanisms have varying frequency dependencies.
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The document provides an overview of principles of electric and electronics, including:
- Basic components of an electrical system including sources, control, loads, and transmission.
- Distinguishing between electromotive force (EMF) and potential difference.
- Defining voltage, current, and units of measurement like volts and amps.
- Describing passive circuit elements like resistors and capacitors, and active elements like batteries.
- Introducing concepts of charge, circuits, and measurement instruments.
Electrophoresis
Electrophoresis is the movement of charged particles through an electrode when subjected to an electric Field
Cations move towards cathode
Anions move towards anode
By this technique solutes are separated by their different rates of travel through an electric field.
Commonly used in biological analysis, particularly in the separations of proteins, peptides and nucleic acids
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ELECTROPHORESIS-mobile.pptx
Electro refers to the energy of electricity and phoresis, means to carry across.
Electrophoresis is a technique for separating charged molecule in a mixture under the influence of an applied electric field.
PAPER ELECTROPHORESIS.pptx
Electrophoresis is a scientific laboratory technique that is used to separate DNA, RNA, or protein molecules based on their size and electrical charge. An electric current is passed through the molecules to move them so that they can be separated via a gel. The pores present in the gel work like a sieve, allowing smaller molecules to pass through more quickly and easily than the larger molecules. According to the way conditions are adjusted during electrophoresis, the molecules can be separated in the desired size range.
What is electrophoresis and what are its uses?
Electrophoresis is a very broadly used technique that, fundamentally, applies electric current to biological molecules – they’re usually DNA, but they can be protein or RNA, too – and separates these fragments into pieces that are larger or smaller in size.
The phenomenon of electrophoresis was first observed by Russian professors Peter Ivanovich Strakhov and Ferdinand Frederic Reuss in 1807 at Moscow University. A constant application of electric field caused the particles of clay dispersed in water to migrate, showing an electrokinetic phenomenon.
Electrophoresis can be defined as an electrokinetic process that separates charged particles in a fluid using an electrical field of charge. Electrophoresis of cations or positively charged ions is sometimes referred to as cataphoresis (or cataphoretic electrophoresis). In contrast, sometimes, the electrophoresis of anions or negatively charged ions is referred to as anaphoresis (or anaphoric electrophoresis).
It’s used in a variety of applications. Though it is most often used in life sciences to separate protein molecules or DNA, it can be achieved through several different techniques and methods depending upon the type and size of the molecules.
The methods differ in some ways, but all we need is a source for the electrical charge, a support medium and a buffer solution. Electrophoresis is also used in laboratories for the separation of molecules based on their size, density and purity.
The method used to separate macromolecules such as DNA, RNA, or protein molecules is known as gel electrophoresis.
It is used in forensics for –
Nucleic acid molecule sizing
DNA fragmentation for southern blotting
RNA fragmentation for northern blotting
Protein fragmentation for western blotting
Separation of PCR products analysis
Detection and analysis of variations or mutations in the sequence
Its clinical applications involve –
Serum protein electrophoresis
Lipoprotein analysis
Diagnosis of haemoglobinopathies and hemoglobin A1c.
The fundamental principle of electrophoresis is the existence of charge separation between the surface of a particle and the fluid immediately surrounding it. An applied electric field acts on the resulting charge density, causing the particle to migrate and the fluid around the particle to flow.
It is the process of separation or purification of protein molecules, DNA, or RNA that differ in charge, size.
Electrophoresis
This presentation provides an overview of electrophoresis. It begins by defining electrophoresis as the differential migration of ionized molecules in an electric field based on their charge-to-mass ratio. It then discusses the basic principles, factors affecting mobility, supporting media like paper and gels, common techniques including low voltage and capillary electrophoresis, and applications for analyzing proteins, DNA, antibiotics and more. The presentation concludes that electrophoresis is a useful method for separating charged substances, from small ions to large molecules, and is widely used despite some limitations.
Rupam sahu electrophoresis ppt
Electrophoresis is a separation Technique that is based on the movement of charged particles in an electric field.
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1. Dielectric materials are insulators in which electrons are tightly bound and there is no free charge. They can be classified as active or passive.
2. Active dielectrics readily store electrical energy, like piezoelectric and ferroelectric materials. Passive dielectrics restrict electrical flow and act as insulators, like glass and rubber.
3. When dielectric materials are placed in an electric field, dipoles are induced through mechanisms like electronic, ionic, orientational and space charge polarization, contributing to the material's dielectric properties. The different polarization mechanisms have varying frequency dependencies.
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The document provides an overview of principles of electric and electronics, including:
- Basic components of an electrical system including sources, control, loads, and transmission.
- Distinguishing between electromotive force (EMF) and potential difference.
- Defining voltage, current, and units of measurement like volts and amps.
- Describing passive circuit elements like resistors and capacitors, and active elements like batteries.
- Introducing concepts of charge, circuits, and measurement instruments.
Electrophoresis
Electrophoresis is the movement of charged particles through an electrode when subjected to an electric Field
Cations move towards cathode
Anions move towards anode
By this technique solutes are separated by their different rates of travel through an electric field.
Commonly used in biological analysis, particularly in the separations of proteins, peptides and nucleic acids
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A student presented their work on cyclic voltammetry. They prepared a 2mM potassium ferricyanide solution with 1M KNO3 as the supporting electrolyte. They polished a platinum working electrode and performed cyclic voltammetry under nitrogen purge. The resulting cyclic voltammogram showed a cathodic peak when Fe(CN)6^3- was reduced and an anodic peak when Fe(CN)6^4- was reoxidized. Analysis of the voltammogram provided information about the redox reaction such as peak potentials and currents. The student demonstrated the experimental procedure for cyclic voltammetry.
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field.
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This document discusses gel electrophoresis, which separates biomolecules like DNA, RNA, and proteins based on their size and charge. It describes the basic principles of electrophoresis, including how mobility depends on factors like net charge and size. The key steps of gel electrophoresis are explained, such as preparing the gel, loading samples, running the current, and staining. Two-dimensional gel electrophoresis is also summarized, which separates proteins based on isoelectric focusing in the first dimension and size in the second dimension, allowing high resolution separation of thousands of proteins.
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This document provides an introduction to fundamentals of electrochemistry, including:
1. Redox reactions involve the transfer of electrons between chemical species and can be monitored by measuring electric current.
2. A redox titration uses the transfer of electrons between analyte and titrant for analytical purposes. Reduction occurs when a substance gains electrons and oxidation occurs when a substance loses electrons.
3. Galvanic cells use spontaneous redox reactions to generate electricity by separating the half-cell reactions and allowing electrons to flow through an external circuit.
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3. The Nernst equation is introduced to calculate cell potentials under non-standard conditions when concentrations vary from standard values. This allows electrochemical cells to be used as probes of chemical equilibria.
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Introduction to Electrophoresis and its Classifications
- 2. DEFINITION It's the movement of charged particles in an electrolyte when subjected to an electric field Uses (Separation of) Serum Urine CSF Biological Fluids
- 3. PRINCIPLE Velocity of movement of particles in an electric field which depends on :- The net charge of molecule, strength of electric field It is inversely proportional to molecular weight
- 4. THEORY Movement of charged molecules in a medium subjected to an electric field v = Eq / f — Equation 1 v = Velocity of molecule E = Electric field (V) q = net charge of molecule F = Frictional coefficient dependent on mass and shape of molecule
- 5. THEORY Electrophoretic mobility (µ): rate of migration per unit field strength µ = v / E — Equation 2 Further derived as µ = Eq / Ef — Equation 3 µ = q / f
- 6. FACTORS AFFECTING ELECTROPHORESIS Net electric charge of the molecule Size and shape of molecule Electric field strength Properties of the supporting medium The pH of medium Temperature of operation Wick flow
- 7. CLASSIFICATION Electrophoresis Technique Position Dimension Zone Moving Horizontal Vertical 1D 2D 3D Boundary Paper Gel Agarose PAGE Capillary
- 8. ZONE ELECTROPHORESIS Technique produces zones which can be observed in a separated manner. Classification Paper Electrophoresis Gel Electrophoresis Agarose Electrophoresis PAGE
- 9. MOVING BOUNDARY ELECTROPHORESIS Technique that's carried out in solution, without a supporting medium Classification Capillary Electrophoresis
- 11. 1D ELECTROPHORESIS • Separation of proteins based on molecular weight over range-10 to 300kDa • Samples weighed and dissolved in Sodium Dodecyl Sulfate (SDS)
- 12. 2D ELECTROPHORESIS Essential tool for separation and fractionation of complex protein mixtures from biological samples Depend on • Isoelectric point (pI) • Isoelectric focusing (IEF)
- 13. 3D ELECTROPHORESIS Protein analysis in a separation medium that extends substantially in 3 spatial dimensions Analyzed in 3 independent separation parameters: Native size Isoelectric Point Molecular weight cc
Editor's Notes
- c