1. Electrophoresis is a technique used to separate charged particles like proteins and nucleic acids based on their size and charge. An electric field is applied, causing the particles to migrate through a medium like paper, gel, or liquid at different rates.
2. Gel electrophoresis involves applying a sample to a gel and running an electric current to separate particles based on size. Agarose and polyacrylamide gels are commonly used media that act as molecular sieves.
3. Paper electrophoresis can separate particles like proteins, but gel electrophoresis provides better resolution. Gel electrophoresis is widely used in research and clinical applications to analyze DNA, proteins, and other biomolecules.
This presentation contain the information about gel electrophoresis method , instruments & types.
Electrophoresis is a method through biological molecules are separated by applying an electric field.
Main purpose of this method is to determine the number , amount & mobility of biological component.
There are some internal & external factors that affects the process of electrophoresis.
The bio-molecules have charge on it & when we apply an electric field , the charge particles move to the opposite cathode. In this way, charge particles are separated
There are 3 types of gels that use in this process .
In this buffers are also used which provide ions that carry a current.
It's my prepared presentation on paper and gel electrophoresis for m.pharm students of 1st year pharmaceutics department.
I hope it will help you well for study.
If you like it then please appreciate it.
Thank you 🤗
Electrophoresis is an electrokinetic process which separates charged particles in a fluid using a field of electrical charge. It is most often used in life sciences to separate protein molecules or DNA and can be achieved through several different procedures depending on the type and size of the molecules. The procedures differ in some ways but all need a source for the electrical charge, a support medium and a buffer solution. Electrophoresis is used in laboratories for the separation of molecules based on size, density and purity.An electric field is applied to molecules and as they are electrically charged themselves it results in a force acting upon them. The greater the charge of the molecule the greater the force applied by the electrical field and therefore the further through the support medium the molecule will move relative to its mass.
Some example applications of electrophoresis include DNA and RNA analysis as well as protein electrophoresis which is a medical procedure used to analyse and separate the molecules found in a fluid sample (most commonly blood and urine samples).Different types of gels are usually used as the support medium for electrophoresis and this may be in slab or tube form depending on which is more beneficial. Gel slabs enable many samples to be run simultaneously and so are frequently used in laboratories. However, tube gels give a better resolution of the results so are often chosen for protein electrophoresis.
Agarose gel is commonly used for electrophoresis of DNA. It has a large pore structure allowing larger molecules to move easily but it is not suitable for sequencing smaller molecules.
Polyacrylamide gel electrophoresis (PAGE) has a clearer resolution than agarose gel making it more suitable for quantitative analysis. This makes it possible to identify how proteins bind to DNA. It can also be used to develop an understanding of how bacteria is becoming resistant to antibiotics through plasmid analysis.
electrophoresis: movement of charge particles in a gel under the influence of an electric field, principle, factors, apparatus, types , application, advantage and disadvantage.
This presentation contain the information about gel electrophoresis method , instruments & types.
Electrophoresis is a method through biological molecules are separated by applying an electric field.
Main purpose of this method is to determine the number , amount & mobility of biological component.
There are some internal & external factors that affects the process of electrophoresis.
The bio-molecules have charge on it & when we apply an electric field , the charge particles move to the opposite cathode. In this way, charge particles are separated
There are 3 types of gels that use in this process .
In this buffers are also used which provide ions that carry a current.
It's my prepared presentation on paper and gel electrophoresis for m.pharm students of 1st year pharmaceutics department.
I hope it will help you well for study.
If you like it then please appreciate it.
Thank you 🤗
Electrophoresis is an electrokinetic process which separates charged particles in a fluid using a field of electrical charge. It is most often used in life sciences to separate protein molecules or DNA and can be achieved through several different procedures depending on the type and size of the molecules. The procedures differ in some ways but all need a source for the electrical charge, a support medium and a buffer solution. Electrophoresis is used in laboratories for the separation of molecules based on size, density and purity.An electric field is applied to molecules and as they are electrically charged themselves it results in a force acting upon them. The greater the charge of the molecule the greater the force applied by the electrical field and therefore the further through the support medium the molecule will move relative to its mass.
Some example applications of electrophoresis include DNA and RNA analysis as well as protein electrophoresis which is a medical procedure used to analyse and separate the molecules found in a fluid sample (most commonly blood and urine samples).Different types of gels are usually used as the support medium for electrophoresis and this may be in slab or tube form depending on which is more beneficial. Gel slabs enable many samples to be run simultaneously and so are frequently used in laboratories. However, tube gels give a better resolution of the results so are often chosen for protein electrophoresis.
Agarose gel is commonly used for electrophoresis of DNA. It has a large pore structure allowing larger molecules to move easily but it is not suitable for sequencing smaller molecules.
Polyacrylamide gel electrophoresis (PAGE) has a clearer resolution than agarose gel making it more suitable for quantitative analysis. This makes it possible to identify how proteins bind to DNA. It can also be used to develop an understanding of how bacteria is becoming resistant to antibiotics through plasmid analysis.
electrophoresis: movement of charge particles in a gel under the influence of an electric field, principle, factors, apparatus, types , application, advantage and disadvantage.
INTRODUCTION, DEFINATION OF ELECTROPHORESIS, ELECTROPHORESIS PRINCIPLE, TYPES OF ELECTROPHORESIS, FREE ELECTROPHORESIS, ZONE ELECTROPHORESIS,PAPER ELECTROPHORESIS, WORKING OF PAPER ELECTROPHORESIS, PROCEDURE FOR PAPER ELECTROPHORESIS, VISUALISATION, FACTORS AFFECTING SEPARATION OF MOLECULES, APPLICATIONS, working of paper electrophoresis ,procedure for paper electrophoresis ,visualisation ,factors affecting separation of molecules ,applications ,forensics ,dna fingerprinting ,molecular biology ,microbiology information about the organisms ,biochemistry mapping of cellular components ,paper electrophoresis is also used in study of sic ,hemoglobin abnormalities ,separation of blood clotting factors ,serum plasma proteins from blood sample ,used in separation and identification of alkaloids ,used for testing water samples ,toxicity of water ,drug industry to determine presence of illelgal drUGS
electrophoresis: types, advantages, disadvantages and applications.Cherry
Electrophoresis is a general term that describes the migration and separation of charged particles under the influence of an electric field.
The particles maybe simple ions, complex macromolecules and colloids or particulate matter- either living cells such as bacteria or inert material such as oil emulsion, droplet etc.
The pores present in the gel work like a sieve, allowing the smaller molecules to pass through more quickly and easily than the larger molecules.
Gel Electrophoresis:
Definition:
Gel electrophoresis is a laboratory technique used to separate and analyze macromolecules such as DNA, RNA, and proteins based on their size and charge. It is a fundamental tool in molecular biology and biochemistry research.
Principle:
The basic principle involves applying an electric field to a gel matrix, typically made of agarose or polyacrylamide. When a current is applied, charged molecules migrate through the gel at rates determined by their size and charge. Smaller molecules move more quickly through the gel, while larger ones move more slowly.
Applications:
Gel electrophoresis is widely used in various applications, including:
-DNA fingerprinting
-Analysis of PCR products
-Checking the purity of nucleic
acid samples
-Protein analysis and
characterization
Honest Reviews of Tim Han LMA Course Program.pptxtimhan337
Personal development courses are widely available today, with each one promising life-changing outcomes. Tim Han’s Life Mastery Achievers (LMA) Course has drawn a lot of interest. In addition to offering my frank assessment of Success Insider’s LMA Course, this piece examines the course’s effects via a variety of Tim Han LMA course reviews and Success Insider comments.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
INTRODUCTION, DEFINATION OF ELECTROPHORESIS, ELECTROPHORESIS PRINCIPLE, TYPES OF ELECTROPHORESIS, FREE ELECTROPHORESIS, ZONE ELECTROPHORESIS,PAPER ELECTROPHORESIS, WORKING OF PAPER ELECTROPHORESIS, PROCEDURE FOR PAPER ELECTROPHORESIS, VISUALISATION, FACTORS AFFECTING SEPARATION OF MOLECULES, APPLICATIONS, working of paper electrophoresis ,procedure for paper electrophoresis ,visualisation ,factors affecting separation of molecules ,applications ,forensics ,dna fingerprinting ,molecular biology ,microbiology information about the organisms ,biochemistry mapping of cellular components ,paper electrophoresis is also used in study of sic ,hemoglobin abnormalities ,separation of blood clotting factors ,serum plasma proteins from blood sample ,used in separation and identification of alkaloids ,used for testing water samples ,toxicity of water ,drug industry to determine presence of illelgal drUGS
electrophoresis: types, advantages, disadvantages and applications.Cherry
Electrophoresis is a general term that describes the migration and separation of charged particles under the influence of an electric field.
The particles maybe simple ions, complex macromolecules and colloids or particulate matter- either living cells such as bacteria or inert material such as oil emulsion, droplet etc.
The pores present in the gel work like a sieve, allowing the smaller molecules to pass through more quickly and easily than the larger molecules.
Gel Electrophoresis:
Definition:
Gel electrophoresis is a laboratory technique used to separate and analyze macromolecules such as DNA, RNA, and proteins based on their size and charge. It is a fundamental tool in molecular biology and biochemistry research.
Principle:
The basic principle involves applying an electric field to a gel matrix, typically made of agarose or polyacrylamide. When a current is applied, charged molecules migrate through the gel at rates determined by their size and charge. Smaller molecules move more quickly through the gel, while larger ones move more slowly.
Applications:
Gel electrophoresis is widely used in various applications, including:
-DNA fingerprinting
-Analysis of PCR products
-Checking the purity of nucleic
acid samples
-Protein analysis and
characterization
Honest Reviews of Tim Han LMA Course Program.pptxtimhan337
Personal development courses are widely available today, with each one promising life-changing outcomes. Tim Han’s Life Mastery Achievers (LMA) Course has drawn a lot of interest. In addition to offering my frank assessment of Success Insider’s LMA Course, this piece examines the course’s effects via a variety of Tim Han LMA course reviews and Success Insider comments.
2024.06.01 Introducing a competency framework for languag learning materials ...Sandy Millin
http://sandymillin.wordpress.com/iateflwebinar2024
Published classroom materials form the basis of syllabuses, drive teacher professional development, and have a potentially huge influence on learners, teachers and education systems. All teachers also create their own materials, whether a few sentences on a blackboard, a highly-structured fully-realised online course, or anything in between. Despite this, the knowledge and skills needed to create effective language learning materials are rarely part of teacher training, and are mostly learnt by trial and error.
Knowledge and skills frameworks, generally called competency frameworks, for ELT teachers, trainers and managers have existed for a few years now. However, until I created one for my MA dissertation, there wasn’t one drawing together what we need to know and do to be able to effectively produce language learning materials.
This webinar will introduce you to my framework, highlighting the key competencies I identified from my research. It will also show how anybody involved in language teaching (any language, not just English!), teacher training, managing schools or developing language learning materials can benefit from using the framework.
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This Gasta posits a strategic approach to integrating AI into HEIs to prepare staff, students and the curriculum for an evolving world and workplace. We will highlight the advantages of working with these technologies beyond the realm of teaching, learning and assessment by considering prompt engineering skills, industry impact, curriculum changes, and the need for staff upskilling. In contrast, not engaging strategically with Generative AI poses risks, including falling behind peers, missed opportunities and failing to ensure our graduates remain employable. The rapid evolution of AI technologies necessitates a proactive and strategic approach if we are to remain relevant.
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2. Introduction
• Electrophoresis may be defined as migration of charged particles through
solution under the influence of an external electric field.
• Ions having positive charge migrate towards a negative electrode and ions
with negative charge migrate toward a positive electrode.
• Ions have different migration rates depending on their total charge, size &
shape. So they can be separated.
• More highly charged ions and ions with smaller size ( higher charge to size
ratio) migrate at faster rate than ions with large size and low charge.
3. • The principle of electrophoresis is when electricity is applied to the medium containing biological
molecules, depending on their net charge & molecular size, they migrate differentially. Thus diff. Proteins/
DNA can be separated .
The force created by electric field can be determined by F = EQ. F is the force, E is electric field & Q is the
charge on molecule or ion.
Thus, more is the charge more is the force on molecules with same size.
4. Types of Electrophoresis
1. Zone Electrophoresis
• Paper electrophoresis
• Gel electrophoresis
• Thin layer electrophoresis
• Cellulose acetate electrophoresis
2. Free Electrophoresis
• Moving boundary electrophoresis
• Capillary electrophoresis • Isoelectric focusing
• Isotachophoresis • Immuno electrophoresis
5. Zone Electrophoresis
• It involves migration of the charged particle on the supporting media.
• Paper, gel, cellulose acetate membrane, starch, poly acrylamide.
• The components that are separated are distributed into discrete zones on
supporting media.
• Supporting media is saturated with buffer solution, small amount of sample
is applied as narrow band .
• Substances separated are nucleic acid, protein, bio polymers .
6. Advantages & Disadvantages
• Easy maintenance & low cost
• Useful in biochemical investigations
• Small amount of sample can be Analyzed
• Unsuitable for accurate mobility and iso electric point determination
• General Method Of Operation :
• Initially Saturation of media with buffer followed by sample application and
electrophoretic separation & then removal of supporting media.
8. 1. Paper Electrophoresis
• In this, sample to be separated is applied to a strip of paper moisturised using a
kind of buffer solution. Eg. Diethyl barbituric acid & barbituric acid dissolved
in alkali, pH 8.6 .
• There are separate tanks of this buffer solution and each end of paper is dipped
in these tanks.
• Different types of cathode and anode are used.
• An electric current is applied, which forces the sample to move on direction of
electrode with opposite polarity.
• Separation takes place in 12 to 14 hours.
9. Principle
• When charged molecules are placed in an electric field, they migrate towards
either the positive or negative electrode according to their charges. In contrast
to proteins, which can have either a net positive or net negative negative
charge, nucleic acids have a consistent negative charge imparted by their
phosphate backbone, & migrate towards anode .
• Equipments : Power pack - It provides a stable direct current & has control for
both voltage & current output.
Electrophoretic cell : It contains electrodes, buffer reservoirs, a support for
paper & a transparent insulating cover. Electrodes are usually made of platinum.
10. Procedure
1. A long strip of filter paper is moistened with a suitable buffer solution of desired pH
& the sample is applied transversely across the central part of strip.
2. Ends are fixed to dip in buffer solution in 2 troughs fitted with electrodes.
3. Electric field of about 20 volts/cm is established.
4. The charged particles of sample migrate along the strip towards respective electrodes
of opposite polarity, according to net charges, sizes & interaction with solid matrix.
5. Homogenous group of particles migrate as separate band.
6. Electrophoresis is carried out for 16 - 18 hours.
7. Proteins are stained ( bromophenol blue ) to make them visible.
8. The separated bands appear as distinct band.
11. 1. Advantages : It is economical.
• It is easy to use.
2. Disadvantages : Certain compounds such as protein, hydrophilic
molecules can’t be resolved due to adsorptive and ionogenic properties of
paper which results in tailing and distortion of components bands.
• Electro osmosis.
Types of Paper Electrophoresis: Horizontal paper Electrophoresis
Vertical Paper Electrophoresis
12.
13. In horizontal paper electrophoresis, the
gel is cast horizontally & placed in
chamber ( filled with running buffer )
that is divided into two sections with
gel in the middle, forming positive
charge at one end & negative at
another.
A continuous running buffer is used
in horizontal gel electrophoresis.
14. Vertical gel electrophoresis contains stacking gel
and resolving gel. The stacking gel concentrates
proteins that are loaded into the well so that the
proteins can start to migrate at the same time. After
stacking, the resolution gel separate proteins based
on the molecular size.
It can give important diagnostic information
concerning serum proteins, and is invaluable
in the differential diagnosis of diseases in
which there are abnormal hemoglobins.
15. Application
1. It is simple, inexpensive & accurate laboratory procedure for various research & clinical
studies.
2. Clinically it is used to study sick cell anaemia, haemoglobin abnormalities & separation of
blood clotting factors & serum plasma protein from blood sample.
3. It is also used in separation & identification of alkaloids.
4. Paper Electrophoresis can also be used for testing water samples, toxicity of water & other
environmental components.
5. Drug - testing industry uses this technique to determine presence of illegal drugs crime
suspects.
6. Separation of amino acids, protein serum & anti biotic .
16. 2. Gel Electrophoresis
• In this, separation is brought about through molecular sieving technique based on
molecular size of substance.
• Gel material acts as as molecular sieve.
• Gel is a colloid in a solid form ( 99% in water ).
• Support media should be electrically neutral.
• Different types of gels which can be used are agar, agarose gel, starch & polyacrylamide
gels ( most common due to high stability).
• The gel acts as a sieve by reducing by movement of macromolecules and allowing micro
molecules to move freely and are used to separate DNA, RNA & Proteins.
17. Agar and Agarose Gel
• Agar is a mixture of polysaccharides extracted from sea weed.
• Agarose is a highly purified uncharged polysaccharide derivative of agar.
• Agarose is chemically basic disaccharide repeating units of 3,6 – Anhydro-L- Galactose.
• Agarose dissolves in boiling liquid. It remains liquid until the temperature lowered to about 40
degree Celsius.
• Pore size may be pre determined by adding the concentration agarose in gel.
• Agarose gels are fragile as they are held together by weak H bonds.
• Pores of agarose gel are large. It is used to seperate macromolecules such as nucleic acid, large
proteins ( albumin ) & protein complex.
18. Polyacrylamide Gel Electrophoresis (PAGE)
• It is prepared by polymerising acrylamide monomer in presence of Methylene- Bis-
Acrylamide to crosslink the monomer ( covalent crosslink)
• Polyacrylamide gel are tougher than agarose gel.
• It is thermostable, transparent, strong & relatively chemically inert.
• PAGE is used in forensic chemistry, biochemistry, genetics, molecular biology &
biotechnology to separate biological macromolecules, usually protein & nucleic acid.
• Protein are separated by molecular sieving ( based on charge to mass ratio and molecular
size)
19. • Advantages: Polyacrylamide gels are stable over wide range of PH &
temperature.
• Simple & separation speed is good enough.
• Types of PAGE : 1. Native PAGE
2. Denatured PAGE or SDS- PAGE
1. Native PAGE : Native gels are run in non denaturing conditions, so that the
analyte natural structure is maintained. Separation is based upon charge, size and
shape of macromolecules.
Useful for separation and purification of mixture of proteins.
2. SDS PAGE : In this separation is based on molecular weight of Proteins.
Useful for checking purity of protein samples.
20. Principle
• It consists in the separation of molecules on the basis of their movement rate
through a gel under the influence of an electrical field. An electric current is
applied across the gel so that one end of the gel has a positive charge and the other
end has a negative charge. The movement of charged molecules is called migration.
Molecules migrate towards the opposite charge.
• Gel electrophoresis is a widely used technique for the analysis of DNA, nucleic acids
and proteins.
21. • Gel electrophoresis separates DNA fragments by size in a solid support medium such as
an agarose gel. Sample (DNA) are pipetted into the sample wells, followed by the
application of an electric current which causes the negatively-charged DNA to migrate
(electrophorese) towards the anodal, positive end. The rate of migration is proportional to
size: smaller fragments move more quickly and wind up at the bottom of the gel.
• DNA is visualised by including in the gel an intercalating dye, ethidium bromide. DNA
fragments take up the dye as they migrate through the gel. Illumination with ultraviolet
light causes the intercalated dye to fluoresce.
• The larger fragments fluoresce more intensely. Although each of the fragments of a
single class of molecule is present in equimolar proportions, the smaller fragments
include less mass of DNA, take up less dye, and therefore fluoresce less intensely. A
“ladder” set of DNA fragments of known size can be run simultaneously and used to
estimate the sizes of the other unknown fragments.
22. Equipments for Gel Electrophoresis
The equipment and supplies necessary for conducting agarose gel electrophoresis are
relatively simple and include:
1.An electrophoresis chamber and power supply
2.Gel casting trays, which are available in a variety of sizes and composed of UV
transparent plastic. The open ends of the trays are closed with tape while the gel is
being cast, then removed prior to electrophoresis.
3.Sample combs, around which molten medium is poured to form sample wells in
the gel.
4.Electrophoresis buffer, usually Tris-acetate-EDTA (TAE) or Tris-borate-EDTA
(TBE).
24. 5. Loading buffer, which contains something dense (e.g. glycerol) to allow
the sample to “fall” into the sample wells, and one or two tracking dyes,
which migrate in the gel and allow visual monitoring or how far the
electrophoresis has proceeded.
6. Staining: DNA molecules are easily visualised under an ultraviolet lamp
when electrphoresed in the presence of the extrinsic fluorescent ethidium
bromide. Alternatively, nucleic acids can be stained after electrophoretic
separation by soaking the gel in a solution of ethidium bromide. When
intercalated into double stranded DNA, fluorescence of this molecule
increases greatly. It is also possible to detect DNA with the extrinsic
fluorescent 8 -anilinonaphthalene -1- sulphonic acid.
7. Transilluminator (an ultraviolet light box), which is used to visualise
stained DNA in gels.
25.
26. Applications
Gel electrophoresis is a routinely used method for separating proteins, DNA or RNA.
• Analysis of PCR products, e.g. in molecular genetic diagnosis or genetic fingerprinting
• Separation of restricted genomic DNA prior to Southern analysis, or of RNA prior to
Northern analysis.
• The agarose gel electrophoresis is widely employed to estimate the size of DNA
fragments after digesting with restriction enzymes, e.g. in restriction mapping of cloned
DNA.
• In the separation of DNA fragments for DNA fingerprinting to investigate crime scenes.
To analyse results of polymerase chain reaction. To analyse genes associated with a
particular illness. In DNA profiling for taxonomy studies to distinguish different species
27. Applications
• Agarose gel electrophoresis is commonly used to resolve circular DNA with
different supercoiling topology, and to resolve fragments that differ due to
DNA synthesis.
• In addition to providing an excellent medium for fragment size analyses,
agarose gels allow purification of DNA fragments. Since purification of
DNA fragments size separated in an agarose gel is necessary for a number
molecular techniques such as cloning, it is vital to be able to purify
fragments of interest from the gel.