It is a subtype of the gel electrophoresis whereby the normal gel is replaced with polyacrylamide gels used as support media.
Gels are made by free radical-induced polymerization of acrylamide and N,N’-Methylenebisacrylamide.
It is the most widely used technique of electrophoresis.
wo-dimensional gel electrophoresis, abbreviated as 2-DE or 2-D electrophoresis, is a form of gel electrophoresis commonly used to analyze proteins. Mixtures of proteins are separated by two properties in two dimensions on 2D gels. 2-DE was first independently introduced by O'Farrell and Klose in 1975.
PAGE is a subtype of the gel electrophoresis whereby the normal gel is replaced with polyacrylamide gels use as the support matrix.
widely used and has very much importance.
COMPLETE PROCEDURE & USES are described in the slide.
It is a subtype of the gel electrophoresis whereby the normal gel is replaced with polyacrylamide gels used as support media.
Gels are made by free radical-induced polymerization of acrylamide and N,N’-Methylenebisacrylamide.
It is the most widely used technique of electrophoresis.
wo-dimensional gel electrophoresis, abbreviated as 2-DE or 2-D electrophoresis, is a form of gel electrophoresis commonly used to analyze proteins. Mixtures of proteins are separated by two properties in two dimensions on 2D gels. 2-DE was first independently introduced by O'Farrell and Klose in 1975.
PAGE is a subtype of the gel electrophoresis whereby the normal gel is replaced with polyacrylamide gels use as the support matrix.
widely used and has very much importance.
COMPLETE PROCEDURE & USES are described in the slide.
Introduction, Principle, Instrumentation and Applications of SDS-PAGEMohammed Mubeen
The following presentation contains helpful information regarding SDS-PAGE, including the history, introduction, principle, instrumentation, advantages and applications of SDS-PAGE.
Gel electrophoresis native, denaturing&reducingLovnish Thakur
Electrophoresis is a technique used to separate and sometimes purify macromolecules - especially proteins and nucleic acids - that differ in size, charge or conformation.
2D-Electrophoresis is an important technique that is being used extensively in the Biochemistry and molecular biology for the quantification of different bio-molecules. It is also used in the different researches like cancer study etc. This presentation covers the introduction, sample preparation, main methodology and steps, staining techniques, applications, cost and availability across Pakistan. It also explains that why there is a need to replace the Electrophoresis with 2D electrophoresis. The main purpose of this effort is to highlight the main points about 2D-Electrophoresis.
A biochemical technique used for isolation of molecules on the basis of size ,shape & density. Categorized into two types:
1) Analytical ultracentrifugation
2) Preparative ultracentrifugation
Centrifugation principle and types by Dr. Anurag YadavDr Anurag Yadav
concept of cnetrifugation,
basic Principle
centrifugal force
types of centrifugation based on use and rotor type
application of the each type of centrifuge
Ultracentrifuge in detail
application in general
Electrophoresis is a separation technique that is based on the movement of charged particles in an electric field
The term electrophoresis was coined from a greek work “Phoresis” which means “Being carried away”
Hence literal meaning of the word electrophoresis means “to carry with electricity.”
an analytical method used to separate components of a protein mixture based on size.
Principle : A charged molecule will migrate in an electric field towards an electrode with opposite sign.
Polyacrylamide gels are characterized by two parameters: total monomer concentration (%T, in g/100 ml) and weight percentage of cross linker (%C).
%T -the relative pore size of the resulting polyacrylamide gel
higher %T - smaller pores.
The practical ranges for monomer concentration are stock solutions of 30-40%
Introduction, Principle, Instrumentation and Applications of SDS-PAGEMohammed Mubeen
The following presentation contains helpful information regarding SDS-PAGE, including the history, introduction, principle, instrumentation, advantages and applications of SDS-PAGE.
Gel electrophoresis native, denaturing&reducingLovnish Thakur
Electrophoresis is a technique used to separate and sometimes purify macromolecules - especially proteins and nucleic acids - that differ in size, charge or conformation.
2D-Electrophoresis is an important technique that is being used extensively in the Biochemistry and molecular biology for the quantification of different bio-molecules. It is also used in the different researches like cancer study etc. This presentation covers the introduction, sample preparation, main methodology and steps, staining techniques, applications, cost and availability across Pakistan. It also explains that why there is a need to replace the Electrophoresis with 2D electrophoresis. The main purpose of this effort is to highlight the main points about 2D-Electrophoresis.
A biochemical technique used for isolation of molecules on the basis of size ,shape & density. Categorized into two types:
1) Analytical ultracentrifugation
2) Preparative ultracentrifugation
Centrifugation principle and types by Dr. Anurag YadavDr Anurag Yadav
concept of cnetrifugation,
basic Principle
centrifugal force
types of centrifugation based on use and rotor type
application of the each type of centrifuge
Ultracentrifuge in detail
application in general
Electrophoresis is a separation technique that is based on the movement of charged particles in an electric field
The term electrophoresis was coined from a greek work “Phoresis” which means “Being carried away”
Hence literal meaning of the word electrophoresis means “to carry with electricity.”
an analytical method used to separate components of a protein mixture based on size.
Principle : A charged molecule will migrate in an electric field towards an electrode with opposite sign.
Polyacrylamide gels are characterized by two parameters: total monomer concentration (%T, in g/100 ml) and weight percentage of cross linker (%C).
%T -the relative pore size of the resulting polyacrylamide gel
higher %T - smaller pores.
The practical ranges for monomer concentration are stock solutions of 30-40%
It is the most common analytical technique used in biochemical estimation in clinical laboratory.
It involves the quantitative estimation of color.
A substance to be estimated colorimetrically, must be colored or it should be capable of forming chromogens (colored complexes) through the addition of reagents.
Back to basics: Fundamental Concepts and Special Considerations in RNA IsolationQIAGEN
RNA integrity and quality are critical to obtain meaningful and reliable downstream data. This slidedeck details the challenges and considerations of handling RNA samples, RNA stabilization, the need for quality control analysis and common methods for RNA integrity and quality assessment.
molecular biology techniques -jaypee university of information technology- ra...RAVI RANJAN
molcular biology techniques- ravi ranjan lb-
contents- basic molecular biology techniques - DNA and RNA isolation from plant sample, nanodrop technique, pcr and cloning.
Electrophoresis along with its history, application and types are discussed here to give a brief yet understanding outlook to the topic explained which is an important topic for the biotechnology as well as all biological research field.
General introduction about electrophoresis
Principle
Working condition of electrophoresis
Factors affecting separation of electrophoresis
Application of electrophoresis
Types of electrophoresis
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
Model Attribute Check Company Auto PropertyCeline George
In Odoo, the multi-company feature allows you to manage multiple companies within a single Odoo database instance. Each company can have its own configurations while still sharing common resources such as products, customers, and suppliers.
How to Split Bills in the Odoo 17 POS ModuleCeline George
Bills have a main role in point of sale procedure. It will help to track sales, handling payments and giving receipts to customers. Bill splitting also has an important role in POS. For example, If some friends come together for dinner and if they want to divide the bill then it is possible by POS bill splitting. This slide will show how to split bills in odoo 17 POS.
Students, digital devices and success - Andreas Schleicher - 27 May 2024..pptxEduSkills OECD
Andreas Schleicher presents at the OECD webinar ‘Digital devices in schools: detrimental distraction or secret to success?’ on 27 May 2024. The presentation was based on findings from PISA 2022 results and the webinar helped launch the PISA in Focus ‘Managing screen time: How to protect and equip students against distraction’ https://www.oecd-ilibrary.org/education/managing-screen-time_7c225af4-en and the OECD Education Policy Perspective ‘Students, digital devices and success’ can be found here - https://oe.cd/il/5yV
Read| The latest issue of The Challenger is here! We are thrilled to announce that our school paper has qualified for the NATIONAL SCHOOLS PRESS CONFERENCE (NSPC) 2024. Thank you for your unwavering support and trust. Dive into the stories that made us stand out!
The Roman Empire A Historical Colossus.pdfkaushalkr1407
The Roman Empire, a vast and enduring power, stands as one of history's most remarkable civilizations, leaving an indelible imprint on the world. It emerged from the Roman Republic, transitioning into an imperial powerhouse under the leadership of Augustus Caesar in 27 BCE. This transformation marked the beginning of an era defined by unprecedented territorial expansion, architectural marvels, and profound cultural influence.
The empire's roots lie in the city of Rome, founded, according to legend, by Romulus in 753 BCE. Over centuries, Rome evolved from a small settlement to a formidable republic, characterized by a complex political system with elected officials and checks on power. However, internal strife, class conflicts, and military ambitions paved the way for the end of the Republic. Julius Caesar’s dictatorship and subsequent assassination in 44 BCE created a power vacuum, leading to a civil war. Octavian, later Augustus, emerged victorious, heralding the Roman Empire’s birth.
Under Augustus, the empire experienced the Pax Romana, a 200-year period of relative peace and stability. Augustus reformed the military, established efficient administrative systems, and initiated grand construction projects. The empire's borders expanded, encompassing territories from Britain to Egypt and from Spain to the Euphrates. Roman legions, renowned for their discipline and engineering prowess, secured and maintained these vast territories, building roads, fortifications, and cities that facilitated control and integration.
The Roman Empire’s society was hierarchical, with a rigid class system. At the top were the patricians, wealthy elites who held significant political power. Below them were the plebeians, free citizens with limited political influence, and the vast numbers of slaves who formed the backbone of the economy. The family unit was central, governed by the paterfamilias, the male head who held absolute authority.
Culturally, the Romans were eclectic, absorbing and adapting elements from the civilizations they encountered, particularly the Greeks. Roman art, literature, and philosophy reflected this synthesis, creating a rich cultural tapestry. Latin, the Roman language, became the lingua franca of the Western world, influencing numerous modern languages.
Roman architecture and engineering achievements were monumental. They perfected the arch, vault, and dome, constructing enduring structures like the Colosseum, Pantheon, and aqueducts. These engineering marvels not only showcased Roman ingenuity but also served practical purposes, from public entertainment to water supply.
2. ELECTROPHORESIS
Electrophoresis is a separation technique that is
based on the movement of charged particles in an
electric field.
The term electrophoresis was coined from a Greek
word “Phoresis” which means “Being Carried
Away”.
Hence literal meaning of the word electrophoresis
means “to carry with electricity.”
3. PRINCIPLE
• Any charged ion or molecule migrates when
placed in an electric field, the rate of migration
depend upon its net charge, size, shape and the
applied electric current.
• Can be represented by following eq.
E*q
V
f
4. PRINCIPLE…CONT.
Whereby,
v = velocity of migration of the molecule.
E = electric field in volts per cm
q = net electric charge on the molecule
f = frictional coefficient
11. GEL TYPES
Polysaccharide extracted
from sea weed.
Gel casted horizontally
Non-toxic.
Separate large molecules
Commonly used for DNA
separations.
Staining can be done
before or pouring the gel.
Cross-linked polymer of
acrylamide.
Gel casted vertically.
Potent neuro-toxic.
Separate small molecules.
Used for DNA or protein
separations.
Staining can be done after
pouring the gel.
Agarose Polyacrylamide Gel
12. POLY ACRYLAMIDE GEL
ELECTROPHORESIS
It is a subtype of the gel electrophoresis whereby
the normal gel is replaced with polyacrylamide gels
used as support media.
Gels are made by free radical-induced
polymerization of acrylamide and N,N‟-
Methylenebisacrylamide.
It is the most widely used technique of
electrophoresis.
15. VISUALIZATION
After the electrophoresis is complete, the molecules in
the gel can be stained to make them visible.
Ethidium bromide, silver, or coomassie blue dye may be
used for this process.
Other methods may also be used to visualize the
separation of the mixture's components on the gel.
If the analyte molecules fluoresce under ultraviolet light, a
photograph can be taken of the gel under ultraviolet
lighting conditions. If the molecules to be separated
contain radioactivity added for visibility, an autoradiogram
can be recorded of the gel.
17. SDS - PAGE
It is a modified version of PAGE whereby Sodium-
dodecyl-sulphate(SDS) is used.
SDS is an amphipathic surfactant.
It denatures proteins by binding to the protein chain with
its hydrocarbon „tail‟, exposing normally buried regions
and „coating‟ the protein chain with surfactant molecules.
The polar „head‟ group of SDS adds an additional benefit
to the use of this denaturant.
19. In their native form, proteins fold into a variety of shapes,
some compact, some elongated.
The rate of migration of native proteins through a sieving
medium is therefore more a reflection of their relative
compactness, and less an accurate measure of molecular
weight.
Denaturing the proteins nullifies structural effects on
mobility, allowing separation on a true charge/mass ratio
basis.
It also separates subunits in multimeric proteins, allowing
analysis of large, complex aggregates.
23. DIFFERENCES
• Separation is based
upon charge, size,
and shape of
macromolecules.
• Useful for
separation and/or
purification of
mixture of proteins
• This was the
original mode of
electrophoresis.
• Separation is based
upon the molecular
weight of proteins.
• The most common
method for
determining MW of
proteins
• Very useful for
checking purity of
protein samples
Native PAGE SDS PAGE
25. APPLICATIONS
Used for estimation of molecular
weight of proteins and nucleic
acids.
Determination of subunit structure
of proteins.
Purification of isolated proteins.
Monitoring changes of protein
content in body fluids.
26. REFERENCES
Indian Pharmacopoeia 2010.
BIOCHEMISTRY by Donald Voet & Judith Voet, Wiley
publications.
BIOCHEMISTRY by Satyanarayana & Chakrapani.
Biochemistry by Upadhyay , Upadhyay and Nath.