This document summarizes the process of agarose gel electrophoresis. Agarose gel is prepared by combining agarose powder with a buffer solution to establish pH and conductivity. Samples are loaded into wells in the gel and an electric current is applied, causing DNA fragments to migrate through the gel at rates corresponding to their size. Agarose gel electrophoresis is used to separate and analyze DNA fragments for applications such as estimating DNA size, analyzing polymerase chain reaction products, and extracting DNA fragments for further purification.
Agarose gel electrophoresis is a technique used to separate DNA fragments by size. DNA samples are loaded into wells in an agarose gel and an electric current is applied, causing the fragments to migrate through the gel at rates dependent on their size. Smaller fragments travel farther than larger fragments, allowing DNA samples to be separated into bands of distinct sizes that can be visualized after staining.
Gel filtration chromatography separates molecules according to size by passing a liquid mobile phase containing the molecules through a column packed with porous beads. Larger molecules are excluded from the pores and elute earlier, while smaller molecules can enter the pores and are retained longer in the column before eluting. It is commonly used to purify and determine the molecular weights of proteins, polymers, and other biomolecules.
1. Ultracentrifugation is a technique that uses very high speeds to separate particles in solution based on properties like size, shape, density.
2. There are two main types - preparative ultracentrifugation which handles large volumes to separate molecules, and analytical ultracentrifugation which uses small volumes and optical detection to study purified molecules.
3. Preparative ultracentrifugation techniques include differential centrifugation, density gradient centrifugation, and zonal centrifugation to separate organelles, proteins, and other molecules. Analytical ultracentrifugation determines molecular weight and detects conformational changes.
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.
Ultracentrifugation is a technique that uses very high rotational speeds, up to 80,000 rpm, to separate particles via centrifugal force up to 600,000g. There are two main types: analytical ultracentrifugation monitors particles in real-time to study molecular interactions and properties, while preparative ultracentrifugation isolates and purifies particles like organelles. Common techniques include differential centrifugation to separate organelles and density gradient centrifugation to separate mixtures based on density.
SDS-PAGE is a technique used to separate proteins by molecular weight. Proteins are denatured and given a negative charge by SDS detergent before running through a polyacrylamide gel matrix by electrophoresis. Smaller proteins migrate faster through the gel, allowing separation by size. After electrophoresis, proteins bands can be visualized using stains like Coomassie blue or silver stain to analyze characteristics like molecular weight, purity, and subunit composition.
Gel-filtration chromatography, also known as size-exclusion chromatography, separates molecules based on their size. Larger molecules pass through the porous gel beads quickly while smaller molecules diffuse into the pores and are delayed. This allows the separation of molecules like proteins, nucleic acids, and polymers by molecular weight. It is a gentle technique that can be used to purify biomolecules without altering buffer conditions.
This document summarizes the process of agarose gel electrophoresis. Agarose gel is prepared by combining agarose powder with a buffer solution to establish pH and conductivity. Samples are loaded into wells in the gel and an electric current is applied, causing DNA fragments to migrate through the gel at rates corresponding to their size. Agarose gel electrophoresis is used to separate and analyze DNA fragments for applications such as estimating DNA size, analyzing polymerase chain reaction products, and extracting DNA fragments for further purification.
Agarose gel electrophoresis is a technique used to separate DNA fragments by size. DNA samples are loaded into wells in an agarose gel and an electric current is applied, causing the fragments to migrate through the gel at rates dependent on their size. Smaller fragments travel farther than larger fragments, allowing DNA samples to be separated into bands of distinct sizes that can be visualized after staining.
Gel filtration chromatography separates molecules according to size by passing a liquid mobile phase containing the molecules through a column packed with porous beads. Larger molecules are excluded from the pores and elute earlier, while smaller molecules can enter the pores and are retained longer in the column before eluting. It is commonly used to purify and determine the molecular weights of proteins, polymers, and other biomolecules.
1. Ultracentrifugation is a technique that uses very high speeds to separate particles in solution based on properties like size, shape, density.
2. There are two main types - preparative ultracentrifugation which handles large volumes to separate molecules, and analytical ultracentrifugation which uses small volumes and optical detection to study purified molecules.
3. Preparative ultracentrifugation techniques include differential centrifugation, density gradient centrifugation, and zonal centrifugation to separate organelles, proteins, and other molecules. Analytical ultracentrifugation determines molecular weight and detects conformational changes.
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.
Ultracentrifugation is a technique that uses very high rotational speeds, up to 80,000 rpm, to separate particles via centrifugal force up to 600,000g. There are two main types: analytical ultracentrifugation monitors particles in real-time to study molecular interactions and properties, while preparative ultracentrifugation isolates and purifies particles like organelles. Common techniques include differential centrifugation to separate organelles and density gradient centrifugation to separate mixtures based on density.
SDS-PAGE is a technique used to separate proteins by molecular weight. Proteins are denatured and given a negative charge by SDS detergent before running through a polyacrylamide gel matrix by electrophoresis. Smaller proteins migrate faster through the gel, allowing separation by size. After electrophoresis, proteins bands can be visualized using stains like Coomassie blue or silver stain to analyze characteristics like molecular weight, purity, and subunit composition.
Gel-filtration chromatography, also known as size-exclusion chromatography, separates molecules based on their size. Larger molecules pass through the porous gel beads quickly while smaller molecules diffuse into the pores and are delayed. This allows the separation of molecules like proteins, nucleic acids, and polymers by molecular weight. It is a gentle technique that can be used to purify biomolecules without altering buffer conditions.
Differential centrifugation is a technique used to separate cellular components like organelles based on sedimentation rate differences caused by varying density, size, shape when spun at increasing centrifugal forces; it involves homogenizing a sample, centrifuging sequentially at low speeds to pellet larger components then higher speeds to further separate smaller ones, allowing fractionation of components from nuclei to ribosomes into pellets and supernatants. Differential centrifugation has applications in separating various mixtures and purifying biomolecules, cells, and subcellular structures.
Gel filtration, also known as size exclusion chromatography, is a technique used to separate biomolecules based on their size. It uses a porous gel or matrix with pores that exclude molecules above a certain size. Small molecules are able to enter the pores and are eluted first, while larger molecules are excluded from the pores and are eluted later. Gel filtration is useful for separating biomolecules without altering pH, ion concentrations, or other conditions. It can be used for desalting, preparative separations, and determining the relative molecular sizes of proteins.
This document provides an overview of electrophoresis. It discusses how electrophoresis works, involving the movement of charged particles through an electrolyte when subjected to an electric field. It then covers the history, factors affecting electrophoresis, types of electrophoresis including paper, SDS-PAGE, native gel, gradient gel, IEF gels, 2D gel electrophoresis, protein blotting, pulsed field gel electrophoresis, and capillary electrophoresis. It also discusses instrumentation, reagents, and applications of various electrophoresis techniques.
This document discusses different types of cell culture, including primary culture, secondary culture, cell lines, and established cell line culture. It describes the process of isolating and culturing cells from tissue samples, including enzymatic and mechanical disaggregation of tissues. The importance of physical environment and culture media for growing cells in vitro is highlighted. Different types of culture media like serum-containing, serum-free, and chemically defined media are also summarized.
This document provides an overview of electrophoresis techniques presented by Miss Sayanti Sau. It discusses the basic principles of electrophoresis and defines different types including zone electrophoresis techniques like paper, gel, thin layer, and cellulose acetate electrophoresis. It also covers moving boundary electrophoresis techniques such as capillary electrophoresis, isotachophoresis, and isoelectric focusing. Details are provided on gel electrophoresis methods including agarose, polyacrylamide, and SDS-PAGE. Applications and advantages of various electrophoresis techniques are highlighted.
This document discusses pulsed field gel electrophoresis (PFGE), a technique used to separate large DNA molecules. It describes how PFGE uses alternating electric fields at different angles to separate DNA fragments from 50kb up to 10Mb in size. Several PFGE system designs are outlined, including Orthogonal-Field Alternation Gel Electrophoresis (OFAGE), Transverse-Alternating Field Gel Electrophoresis (TAFE), Field Inversion Gel Electrophoresis (FIGE), Rotating Gel Electrophoresis (RGE), and Contour-Clamped Homogeneous Electric Fields (CHEF). The document also reviews running conditions and applications of PFGE such as genome mapping, fingerprinting, and studying radiation damage.
Ultracentrifugation is a technique that uses high centrifugal forces generated by rotational speeds of up to 150,000 rpm to separate particles in solution based on differences in size, shape, density, and viscosity. It is an important tool in biochemical research used to isolate molecules like DNA, RNA, lipids, and separate organelles from cells. There are two main types - analytical ultracentrifugation which studies molecular interactions and properties, and preparative ultracentrifugation which isolates and purifies particles using techniques like density gradient centrifugation. Proper rotor selection and maintenance of the centrifuge are important for safe and effective use of this technique.
Autoradiography is a bioanalytical technique that is used to visualize the radioactively labelled substances or molecules or or fragments of molecules by using X-ray films or photographic emulsions.
Standard test that used to determine the charged molecules, mainly proteins and nucleic acids.
Widely used in biochemistry, forensics, genetics and molecular biology.
Laemmli system of SDS-PAGE was first introduced in 1970s
Isoelectric focusing is a technique that separates molecules like proteins based on their isoelectric point, which is the pH at which the molecule has no net charge. It was developed in the 1960s and allows for much better resolution than older techniques. The process involves creating an immobilized pH gradient using carrier ampholytes, loading protein samples, and applying an electric field to cause proteins to migrate to the point in the gradient matching their isoelectric point. The separated proteins can then be visualized through staining. Isoelectric focusing is useful for applications like identifying serum proteins and aiding in proteomics research.
Agarose gel electrophoresis is a method to separate DNA fragments by size using an agarose gel matrix and electric current. Shorter DNA fragments migrate faster and farther than longer ones. DNA is visualized by staining with ethidium bromide and viewing under UV light. Agarose concentration determines resolution, with 0.8% gels best for separating large 5-10kb fragments and 2% for small 0.2-1kb fragments. Applications include estimating DNA size, analyzing PCR products, and separating DNA for further analysis.
Phase-contrast microscopy is a technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image, allowing living cells that are otherwise invisible to be seen. It works by separating light rays that pass through a specimen unchanged from those that are diffracted, using an annular diaphragm and phase plate in the light path. Phase-contrast microscopy is widely used in biological research for observing living cells, microorganisms, and other transparent specimens without staining or fixing.
This document summarizes a seminar presentation on 2D electrophoresis. 2D electrophoresis is a technique used to separate mixed proteins based on their isoelectric point and mass. It involves two sequential electrophoretic steps: iso-electric focusing to separate proteins by charge, followed by SDS-PAGE to separate by molecular weight. The document describes the principles, methods, applications and references for 2D electrophoresis.
This document discusses biological databases and nucleic acid sequence databases. It describes the three primary nucleotide sequence databases: GenBank, EMBL, and DDBJ. GenBank is hosted by the National Center for Biotechnology Information and contains over 286 million bases and 352,000 sequences. EMBL is hosted by the European Molecular Biology Laboratory and mirrors data daily with GenBank and DDBJ. DDBJ is the DNA Data Bank of Japan and also mirrors data daily with the other two databases. Biological databases are important tools for scientists to understand biology at multiple levels.
Reverse phase chromatography is a technique where the binding of solutes to a hydrophobic stationary phase occurs via hydrophobic interactions. It uses a stationary phase with hydrophobic ligands chemically bonded to a solid support. Polar mobile phases are used to elute retained solutes from the column. Key parameters that affect separation include the pH, ionic strength, and polarity of the mobile phase, use of gradients or isocratic elution, column length, and addition of ion-pairing agents. Reverse phase chromatography is commonly used to purify biomolecules like proteins, peptides, and nucleic acids.
Gel electrophoresis is a technique used to separate charged molecules like proteins and nucleic acids based on their size and charge. It involves placing the molecules in an agarose, starch or polyacrylamide gel and applying an electric current, causing the molecules to migrate through the gel at different rates. SDS-PAGE is a common type of gel electrophoresis where sodium dodecyl sulfate is used to denature proteins, allowing separation based on molecular weight. The separated molecules can then be visualized by staining techniques to analyze proteins, detect mutations, and more.
Isoelectric focusing is a technique that separates proteins and other amphoteric molecules based on their isoelectric point, which is the pH at which they have no net charge. During isoelectric focusing, proteins are placed in an immobilized pH gradient formed by carrier ampholytes and an electric field is applied, causing the proteins to migrate through the pH gradient until they reach the pH that matches their isoelectric point and stop migrating. It is useful for analyzing protein heterogeneity and separating isoforms that have small charge differences but similar molecular weights.
Electrophoresis is a technique used to separate molecules like DNA, RNA, and proteins based on their size, charge, and shape. It works by placing the molecules in an electrically charged gel and applying a voltage so that they migrate at different speeds according to their physical properties. Agarose gel electrophoresis is commonly used to separate DNA fragments, while polyacrylamide gel electrophoresis (PAGE) with sodium dodecyl sulfate (SDS) is used to separate denatured proteins by size. Electrophoresis has many applications including DNA analysis, vaccine development, determining antibiotic resistance, and diagnosing diseases.
Differential centrifugation is a technique used to separate cellular components like organelles based on sedimentation rate differences caused by varying density, size, shape when spun at increasing centrifugal forces; it involves homogenizing a sample, centrifuging sequentially at low speeds to pellet larger components then higher speeds to further separate smaller ones, allowing fractionation of components from nuclei to ribosomes into pellets and supernatants. Differential centrifugation has applications in separating various mixtures and purifying biomolecules, cells, and subcellular structures.
Gel filtration, also known as size exclusion chromatography, is a technique used to separate biomolecules based on their size. It uses a porous gel or matrix with pores that exclude molecules above a certain size. Small molecules are able to enter the pores and are eluted first, while larger molecules are excluded from the pores and are eluted later. Gel filtration is useful for separating biomolecules without altering pH, ion concentrations, or other conditions. It can be used for desalting, preparative separations, and determining the relative molecular sizes of proteins.
This document provides an overview of electrophoresis. It discusses how electrophoresis works, involving the movement of charged particles through an electrolyte when subjected to an electric field. It then covers the history, factors affecting electrophoresis, types of electrophoresis including paper, SDS-PAGE, native gel, gradient gel, IEF gels, 2D gel electrophoresis, protein blotting, pulsed field gel electrophoresis, and capillary electrophoresis. It also discusses instrumentation, reagents, and applications of various electrophoresis techniques.
This document discusses different types of cell culture, including primary culture, secondary culture, cell lines, and established cell line culture. It describes the process of isolating and culturing cells from tissue samples, including enzymatic and mechanical disaggregation of tissues. The importance of physical environment and culture media for growing cells in vitro is highlighted. Different types of culture media like serum-containing, serum-free, and chemically defined media are also summarized.
This document provides an overview of electrophoresis techniques presented by Miss Sayanti Sau. It discusses the basic principles of electrophoresis and defines different types including zone electrophoresis techniques like paper, gel, thin layer, and cellulose acetate electrophoresis. It also covers moving boundary electrophoresis techniques such as capillary electrophoresis, isotachophoresis, and isoelectric focusing. Details are provided on gel electrophoresis methods including agarose, polyacrylamide, and SDS-PAGE. Applications and advantages of various electrophoresis techniques are highlighted.
This document discusses pulsed field gel electrophoresis (PFGE), a technique used to separate large DNA molecules. It describes how PFGE uses alternating electric fields at different angles to separate DNA fragments from 50kb up to 10Mb in size. Several PFGE system designs are outlined, including Orthogonal-Field Alternation Gel Electrophoresis (OFAGE), Transverse-Alternating Field Gel Electrophoresis (TAFE), Field Inversion Gel Electrophoresis (FIGE), Rotating Gel Electrophoresis (RGE), and Contour-Clamped Homogeneous Electric Fields (CHEF). The document also reviews running conditions and applications of PFGE such as genome mapping, fingerprinting, and studying radiation damage.
Ultracentrifugation is a technique that uses high centrifugal forces generated by rotational speeds of up to 150,000 rpm to separate particles in solution based on differences in size, shape, density, and viscosity. It is an important tool in biochemical research used to isolate molecules like DNA, RNA, lipids, and separate organelles from cells. There are two main types - analytical ultracentrifugation which studies molecular interactions and properties, and preparative ultracentrifugation which isolates and purifies particles using techniques like density gradient centrifugation. Proper rotor selection and maintenance of the centrifuge are important for safe and effective use of this technique.
Autoradiography is a bioanalytical technique that is used to visualize the radioactively labelled substances or molecules or or fragments of molecules by using X-ray films or photographic emulsions.
Standard test that used to determine the charged molecules, mainly proteins and nucleic acids.
Widely used in biochemistry, forensics, genetics and molecular biology.
Laemmli system of SDS-PAGE was first introduced in 1970s
Isoelectric focusing is a technique that separates molecules like proteins based on their isoelectric point, which is the pH at which the molecule has no net charge. It was developed in the 1960s and allows for much better resolution than older techniques. The process involves creating an immobilized pH gradient using carrier ampholytes, loading protein samples, and applying an electric field to cause proteins to migrate to the point in the gradient matching their isoelectric point. The separated proteins can then be visualized through staining. Isoelectric focusing is useful for applications like identifying serum proteins and aiding in proteomics research.
Agarose gel electrophoresis is a method to separate DNA fragments by size using an agarose gel matrix and electric current. Shorter DNA fragments migrate faster and farther than longer ones. DNA is visualized by staining with ethidium bromide and viewing under UV light. Agarose concentration determines resolution, with 0.8% gels best for separating large 5-10kb fragments and 2% for small 0.2-1kb fragments. Applications include estimating DNA size, analyzing PCR products, and separating DNA for further analysis.
Phase-contrast microscopy is a technique that converts phase shifts in light passing through a transparent specimen to brightness changes in the image, allowing living cells that are otherwise invisible to be seen. It works by separating light rays that pass through a specimen unchanged from those that are diffracted, using an annular diaphragm and phase plate in the light path. Phase-contrast microscopy is widely used in biological research for observing living cells, microorganisms, and other transparent specimens without staining or fixing.
This document summarizes a seminar presentation on 2D electrophoresis. 2D electrophoresis is a technique used to separate mixed proteins based on their isoelectric point and mass. It involves two sequential electrophoretic steps: iso-electric focusing to separate proteins by charge, followed by SDS-PAGE to separate by molecular weight. The document describes the principles, methods, applications and references for 2D electrophoresis.
This document discusses biological databases and nucleic acid sequence databases. It describes the three primary nucleotide sequence databases: GenBank, EMBL, and DDBJ. GenBank is hosted by the National Center for Biotechnology Information and contains over 286 million bases and 352,000 sequences. EMBL is hosted by the European Molecular Biology Laboratory and mirrors data daily with GenBank and DDBJ. DDBJ is the DNA Data Bank of Japan and also mirrors data daily with the other two databases. Biological databases are important tools for scientists to understand biology at multiple levels.
Reverse phase chromatography is a technique where the binding of solutes to a hydrophobic stationary phase occurs via hydrophobic interactions. It uses a stationary phase with hydrophobic ligands chemically bonded to a solid support. Polar mobile phases are used to elute retained solutes from the column. Key parameters that affect separation include the pH, ionic strength, and polarity of the mobile phase, use of gradients or isocratic elution, column length, and addition of ion-pairing agents. Reverse phase chromatography is commonly used to purify biomolecules like proteins, peptides, and nucleic acids.
Gel electrophoresis is a technique used to separate charged molecules like proteins and nucleic acids based on their size and charge. It involves placing the molecules in an agarose, starch or polyacrylamide gel and applying an electric current, causing the molecules to migrate through the gel at different rates. SDS-PAGE is a common type of gel electrophoresis where sodium dodecyl sulfate is used to denature proteins, allowing separation based on molecular weight. The separated molecules can then be visualized by staining techniques to analyze proteins, detect mutations, and more.
Isoelectric focusing is a technique that separates proteins and other amphoteric molecules based on their isoelectric point, which is the pH at which they have no net charge. During isoelectric focusing, proteins are placed in an immobilized pH gradient formed by carrier ampholytes and an electric field is applied, causing the proteins to migrate through the pH gradient until they reach the pH that matches their isoelectric point and stop migrating. It is useful for analyzing protein heterogeneity and separating isoforms that have small charge differences but similar molecular weights.
Electrophoresis is a technique used to separate molecules like DNA, RNA, and proteins based on their size, charge, and shape. It works by placing the molecules in an electrically charged gel and applying a voltage so that they migrate at different speeds according to their physical properties. Agarose gel electrophoresis is commonly used to separate DNA fragments, while polyacrylamide gel electrophoresis (PAGE) with sodium dodecyl sulfate (SDS) is used to separate denatured proteins by size. Electrophoresis has many applications including DNA analysis, vaccine development, determining antibiotic resistance, and diagnosing diseases.
Electrophoresis is a laboratory technique used to separate DNA, RNA, or protein molecules based on their size and electrical charge.
Different types of electrophoresis.
Gel electrophoresis; Agarose Gel electrophoresis; polyacrylamide gel electrophoresis; pulsed-field gel electrophoresis
Electrophoresis is a technique used to separate biomolecules like DNA, RNA, or proteins based on their size and charge. It works by applying an electric current to move the molecules through a gel or medium. Smaller molecules move faster through the pores in the gel than larger molecules, allowing separation. There are different types of electrophoresis that use different gel materials like agarose or polyacrylamide gels and techniques like pulsed field gel electrophoresis to separate different sized molecules. Electrophoresis is widely used in areas like molecular biology, genetics, and clinical testing.
Electrophoresis and its types and its importance in Genetic engineeringSwaatiSharma2
This document discusses electrophoresis, which is a technique used to separate biomolecules like DNA, RNA, and proteins based on their size and charge. It moves through two common media - agarose gel and polyacrylamide gel. Agarose gel is used for larger molecules while polyacrylamide gel provides better resolution for smaller molecules. The document explains the principles, factors affecting separation, different types like agarose gel electrophoresis and polyacrylamide gel electrophoresis. It also discusses the requirements, steps involved and applications of agarose and polyacrylamide gel electrophoresis.
Gel electrophoresis is a technique used to separate biomolecules like DNA, RNA, and proteins based on their size and charge. It works by applying an electric current to move the molecules through a gel, where smaller molecules move faster than larger ones. The document discusses the different types of gels used like agarose, polyacrylamide, and starch gels. It also explains the basic process which involves loading samples into wells, applying a current, and staining bands to visualize the separated biomolecules. Key applications are determining molecular weights and separating DNA or protein fragments.
Electrophoresis is a technique used to separate charged molecules like proteins and nucleic acids based on their size and charge. It works by applying an electric current to move these molecules through a medium like a gel or paper. The document discusses different types of electrophoresis like gel electrophoresis, paper electrophoresis, and isolectric focusing. It also explains how various factors like the molecule's charge, size, and shape affect its movement during electrophoresis.
Gel electrophoresis is a method to separate macromolecules like DNA, RNA, and proteins based on their size and charge by applying an electric current through a gel matrix. Most molecular biology labs use agarose gel electrophoresis to separate and analyze DNA fragments. During electrophoresis, DNA molecules are separated based on their rate of migration through the agarose gel under an applied electric field with smaller fragments moving faster.
This document discusses various types of electrophoresis techniques. It begins by explaining that electrophoresis involves charged molecules like proteins and nucleic acids migrating in response to an electrical field. It then discusses different gel types like agarose and polyacrylamide gels that are used as supporting matrices. It also covers techniques like SDS-PAGE, isoelectric focusing, two-dimensional gel electrophoresis, and capillary electrophoresis. The document provides details on how each technique separates molecules based on properties like size, charge, and isoelectric point.
This document provides information about electrophoresis, including:
- The general principle that charged molecules will migrate toward the electrode of opposite charge in an electrical field.
- Factors that affect electrophoresis like molecular charge, size, shape, strength of electrical field, and temperature.
- Types of electrophoresis including gel electrophoresis where molecules are separated in a gel matrix based on size and charge.
- Applications like analyzing proteins, nucleic acids, and using agarose gel electrophoresis to visualize DNA, RNA, and PCR products.
Electrophoresis is a method used to separate charged molecules like DNA and RNA using an electric field. In agarose gel electrophoresis, DNA or RNA samples are placed into wells in an agarose gel and an electric current is applied, causing the charged molecules to migrate through the gel at rates dependent on their size. Smaller molecules migrate faster through the gel pores than larger molecules, resulting in size-based separation. Factors like agarose concentration, voltage, buffer composition influence separation resolution. Agarose gel electrophoresis is commonly used to analyze and separate DNA fragments between 200 bp to 20 kb in size.
Gel electrophoresis is a method to separate molecules like DNA, RNA, or proteins based on their size and charge. It works by placing the molecules in an agarose or polyacrylamide gel and applying an electric current, causing the molecules to migrate through the gel at different rates depending on their size and charge. Smaller and more highly charged molecules move faster. This allows separation of molecules that differ in size by as little as a single nucleotide or amino acid. Common applications include analyzing DNA fragments after enzymatic digestion or PCR, separating proteins by molecular weight, and determining purity of samples.
Running of Agarose Gel Electrophoresis Practical Sabahat Ali
Electrophoresis technique used for separation of Macromolecules(DNA, Proteins & their derivatives)
Separation occur on the basis of charge to size ratio
Electrophoresis is a technique used to separate charged molecules like proteins and nucleic acids. It works by applying an electric field to migrate these molecules through a buffer or gel based on their size and charge. This document discusses different electrophoresis techniques including paper, gel, capillary electrophoresis and isoelectric focusing. It provides details on how each technique works, advantages and applications.
B.Sc Biotech II BAT Unit 3 ElectrophoresisRai University
Gel electrophoresis is a method used to separate macromolecules like DNA, RNA, and proteins based on their size and charge. It works by applying an electric field to move charged particles through a gel matrix. Smaller molecules move faster and migrate farther than larger ones. Agarose gels are commonly used to separate DNA fragments between 50-20,000 base pairs, while polyacrylamide gels can separate smaller fragments and proteins. The procedure involves casting the gel, loading samples into wells, applying a current to separate the molecules, and visualizing the results.
Agarose gel electrophoresis by KK Sahu sirKAUSHAL SAHU
INTRODUCTION.
HISTORY.
PROCESS OF GEL ELECTROPHORESIS.
AGAROSE GEL ELECTROFORESIS.
POLYACRYALAMIDE GEL ELECTRIPHORESIS.
GEL CONDITION.
DENATURETION.
NATIVE.
BUFFERS.
USES.
CONCLUSION.
REFFERENCES.
Electrophoresis is a technique used to separate charged particles such as proteins or nucleic acids. It involves applying an electric field to migrate these particles through a buffer or gel based on their size and charge. There are several types of electrophoresis including paper, gel, capillary, and moving boundary which utilize different supporting media and techniques to achieve high resolution separations. Electrophoresis is widely used in biochemistry and molecular biology for analytical purposes.
Gel Electrophoresis, ITS FACTOR AFFECTING, ITS TYPES,NORMAL METHODOLOGY, PAGERitamMukherjee11
ELECTROPHORESIS may be defined as the migration of the charged particle through a solution under the influence of an external electric field.
GEL ELECTROPHORESIS is a technique used to separate biomacromolecules (such as DNA, RNA, proteins, etc.) and their fragments based on their size and charge, by applying an electric field to a gel with small pores.
INTRODUCTION - Separation is brought about through molecular sieving techniques, based on the molecular size of the substances. Gel material acts as a “ molecular sieve”
It is important that support media is electrically neutral.
There are different types of gel that can be used, they are Agarose, Polyacrylamide, Starch, and Sephadex.
A porous gel acts as a sieve by retarding or, in some cases by completely obstructing the movement of macromolecules which allowed smaller molecules to migrate freely.
PRINCIPLE - According to charge: When charged molecules are placed in an electric field, they migrate toward either the positive (anode) or negative (cathode) pole according to their charge.
According to size: The smaller molecules move more swiftly than the larger-sized ones, as they can travel through the pores more easily than the later.
According to Molecular weight: The smaller molecular weight will move faster than the larger molecular weight compound.
Production of live food (Aquatic micro animals)for the rearing of fish fry at...Hafiz M Waseem
Production of live food (Aquatic micro animals)for the rearing of fish fry at hatcheries ROTIFERSBrachionus sp. CRUSTACEANS (CLADOCERANS)Moina sp. Daphnia sp.
green water production at fish hatcheries and its uses to enhance primary pro...Hafiz M Waseem
green water production at fish hatcheries and its uses to enhance primary productivity.ppt
Chlorella sp.
Scenedesmus sp.
Tetraselmis chuii
Skeletonemia sp
Spirulina sp.
Chaetoceros sp.
Nitzschia sp.
The biofloc is a protein-rich aggregate of organic material and microorganisms that forms in aquaculture systems. Biofloc technology maintains water quality and provides nutrients by balancing carbon and nitrogen through the addition of carbon sources like molasses. It has been successfully used in tilapia and shrimp farming and allows for high stocking densities through natural water treatment. Key factors that must be controlled include carbon to nitrogen ratio, dissolved oxygen, pH, and ammonia, nitrite and nitrate levels.
Determination of p h of waste water sample .....................................Hafiz M Waseem
ecologyDetermination of pH of Waste Water Sample ..................................................... 4
Determination Dissolved Oxygen within Water ................................................... 5
Adaptive Features of Animals in Relation to Food and Environment .................. 7
Study the Plant Population Density ................................................................... 10
Experimental Design and Approaches to Ecological Research ........................ 12
The document discusses the benefits of exercise for mental health. Regular physical activity can help reduce anxiety and depression and improve mood and cognitive functioning. Exercise causes chemical changes in the brain that may help protect against mental illness and improve symptoms.
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
Trophic levels and energy variation with increasing trophic levels.food chain...Hafiz M Waseem
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
This document contains personal information for Hafiz Muhammad Waseem of Lahore, Pakistan who is taking the course Applied Ecology (ZOOL3118) at the University of Education Lahore, Pakistan. It also lists Books for reference.
I AM HAFIZ MUHAMMAD WASEEM from mailsi vehari
BSc from science college Multan
MSC university of education Lahore
i love Pakistan and my teachers and my parents
How to Setup Warehouse & Location in Odoo 17 InventoryCeline George
In this slide, we'll explore how to set up warehouses and locations in Odoo 17 Inventory. This will help us manage our stock effectively, track inventory levels, and streamline warehouse operations.
How to Make a Field Mandatory in Odoo 17Celine George
In Odoo, making a field required can be done through both Python code and XML views. When you set the required attribute to True in Python code, it makes the field required across all views where it's used. Conversely, when you set the required attribute in XML views, it makes the field required only in the context of that particular view.
How to Build a Module in Odoo 17 Using the Scaffold MethodCeline George
Odoo provides an option for creating a module by using a single line command. By using this command the user can make a whole structure of a module. It is very easy for a beginner to make a module. There is no need to make each file manually. This slide will show how to create a module using the scaffold method.
This presentation includes basic of PCOS their pathology and treatment and also Ayurveda correlation of PCOS and Ayurvedic line of treatment mentioned in classics.
Exploiting Artificial Intelligence for Empowering Researchers and Faculty, In...Dr. Vinod Kumar Kanvaria
Exploiting Artificial Intelligence for Empowering Researchers and Faculty,
International FDP on Fundamentals of Research in Social Sciences
at Integral University, Lucknow, 06.06.2024
By Dr. Vinod Kumar Kanvaria
This presentation was provided by Steph Pollock of The American Psychological Association’s Journals Program, and Damita Snow, of The American Society of Civil Engineers (ASCE), for the initial session of NISO's 2024 Training Series "DEIA in the Scholarly Landscape." Session One: 'Setting Expectations: a DEIA Primer,' was held June 6, 2024.
LAND USE LAND COVER AND NDVI OF MIRZAPUR DISTRICT, UPRAHUL
This Dissertation explores the particular circumstances of Mirzapur, a region located in the
core of India. Mirzapur, with its varied terrains and abundant biodiversity, offers an optimal
environment for investigating the changes in vegetation cover dynamics. Our study utilizes
advanced technologies such as GIS (Geographic Information Systems) and Remote sensing to
analyze the transformations that have taken place over the course of a decade.
The complex relationship between human activities and the environment has been the focus
of extensive research and worry. As the global community grapples with swift urbanization,
population expansion, and economic progress, the effects on natural ecosystems are becoming
more evident. A crucial element of this impact is the alteration of vegetation cover, which plays a
significant role in maintaining the ecological equilibrium of our planet.Land serves as the foundation for all human activities and provides the necessary materials for
these activities. As the most crucial natural resource, its utilization by humans results in different
'Land uses,' which are determined by both human activities and the physical characteristics of the
land.
The utilization of land is impacted by human needs and environmental factors. In countries
like India, rapid population growth and the emphasis on extensive resource exploitation can lead
to significant land degradation, adversely affecting the region's land cover.
Therefore, human intervention has significantly influenced land use patterns over many
centuries, evolving its structure over time and space. In the present era, these changes have
accelerated due to factors such as agriculture and urbanization. Information regarding land use and
cover is essential for various planning and management tasks related to the Earth's surface,
providing crucial environmental data for scientific, resource management, policy purposes, and
diverse human activities.
Accurate understanding of land use and cover is imperative for the development planning
of any area. Consequently, a wide range of professionals, including earth system scientists, land
and water managers, and urban planners, are interested in obtaining data on land use and cover
changes, conversion trends, and other related patterns. The spatial dimensions of land use and
cover support policymakers and scientists in making well-informed decisions, as alterations in
these patterns indicate shifts in economic and social conditions. Monitoring such changes with the
help of Advanced technologies like Remote Sensing and Geographic Information Systems is
crucial for coordinated efforts across different administrative levels. Advanced technologies like
Remote Sensing and Geographic Information Systems
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Changes in vegetation cover refer to variations in the distribution, composition, and overall
structure of plant communities across different temporal and spatial scales. These changes can
occur natural.
3. Introduction
Gel electrophoresis?
Agarose is a polysaccharide
made from seaweed
It is dissolved in buffer,
heated and cooled to a
gelatinous solid in the form
of inert matrix
A gel is a colloid, suspension
of tiny particles in a medium,
occurring in a solid form (like
gelatin)
Gel electrophoresis refers to
the separation of charged
molecules like nucleic acids,
proteins, etc. when an electric
current is applied
It is the easiest analyzing
macromolecules
4. Introduction
How GE works?
Frictional force of the gel
matrix acts as a molecular
sieve which separate the
molecules on the basis of their
size
Macromolecules are forced to
move through the pores when
electrical current is applied
Most agarose gels are made
between 0.7% and 2% of
agarose in some suitable
buffer
The rate of migration through
the electric field depends on
• Charge of the molecules
• Size of the molecules
• Shape of the molecules
5. Introduction
How GE works?
After staining, the separated
macromolecules in the gel
are seen in the form of bands
A standard is also run in one
lane
It is most commonly used
technique in biochemistry
and molecular biology
6. Introduction
Types of GE
Polyacrylamide gel
electrophoresis (PAGE) is used
for separation of proteins
ranging in size from 5 to
2,000 kDa
It uses sodium dodecyl sulfate
(SDS) as detergent which
denature the proteins and
enable them to move
independently of each other
Agarose gel electrophoresis
uses agarose
Pores of an agarose gel are
large so it is used to separate
macromolecules such as
• Nucleic acids
• Large proteins
• Protein complexes
8. Most agarose gels are made
between 0.7% and 3%
Low percentage gels are very
fragile and are used to
separate DNA molecules from
5 to 10 kb in size
While high percentage gels
are used to separate smaller
molecules
DNA and RNA molecules
(negatively charged) move
towards anode when electric
current is applied
Separation depends upon
composition and ionic
strength of buffers used
Properties of AGE
9. Higher the voltage used,
faster the nucleic acids will
separate
Visualization of the nucleic
acids may be achieved by
ethidium bromide, silver
staining, etc.
3-D structure of agarose gels
is held together with
hydrogen bonds
DNA bands can be cut from
the gel and processed further
for the research
Buffers used contain EDTA to
inactivate many nucleases
11. Requirements of AGE
Gel Apparatus
A power supply, a gel
chamber, combs,
micropipettes, ladder,
buffers, dyes, UV illuminator,
electrodes, cables, gel
mixtures, gel doc system, are
needed
12. Requirements of AGE
Buffers
Buffers maintain a pH either
by absorbing or releasing
Hydrogen ions
Buffers - either TBE or TAE
provide ions to ensure
electrical conductivity along
with other functions
Not only agarose is dissolved
in the buffers but the gel slab
is also submerged in the
buffers after solidifying
TAE has the minimum
buffering capacity but has the
best resolution as compared
with other buffers
13. Requirements of AGE
DNA Ladder
One of the wells in the gel is
loaded with a DNA ladder
This is used as a marker to
compare the separated DNA
fragments in the form of
bands in the gel and to
estimate their sizes
14. Requirements of AGE
Visualizing DNA
Ethidium bromide (EtBr), a
fluorescent dye which is
visualized when excited by
UV light is generally used
Gel is soaked in a solution of
EtBr and the DNA bands take
up the dye
Then the gel is placed under
UV illuminator and visualized
and photographed for
further analysis
16. Gel is prepared by dissolving
the agarose powder as per
requirement in the suitable
buffer
It is heated so that agarose is
melted in the buffer
The melted agarose is allowed
to cool before pouring the
solution into a cast
A comb is placed in the cast to
create wells for loading the
samples
Place the gel at 4C or at room
temperature till it is
completely solidified
Procedure of AGE
Casting of gel
17. Once the gel is solidified, the
comb is removed for loading
DNA samples
Loading buffer is mixed with
the samples to increase
density and to add some dye
(bromophenol blue) to the
samples
The gel is placed in
electrophoresis unit and is
covered with the buffer
Load ladder and samples in
the wells
Electrophoresis is carried at
the suitable voltage
Procedure of AGE
Loading of samples
18. Electrophoresis is carried out
horizontally or vertically
Buffer used in the gel is the
same as the running buffer in
the electrophoretic tank
Run the gel at 80-150 volatge
until the dye is about 75-80%
down the gel
Afterwards, the gel is
removed from the gel box
DNA fragments are visualized
on placing the gel under UV
source
Size of DNA fragments is
estimated with the help of
ladder
Procedure of AGE
Electrophoresis
20. It is used to analyze DNA
molecules which are cut by
restriction enzymes
Cut fragments can be used
for cloning purposes
Analysis of PCR products, e.g.
in molecular diagnostics
or genetic fingerprinting
Separation of DNA fragments
for extraction and
purification purposes
Separation of restricted
genomic DNA prior
to Southern transfer or of
RNA before Northern
transfer
Applications of AGE
21. DNA molecules can be easily
recovered from the gels
without any damage
It is also used for screening
protein abnormalities in
various biological fluids like
serum, urine, CSF, etc.
Size of DNA molecules can
be analyzed by lambda DNA
ladder
Two dimensional
electrophoresis is useful for
identifying a particular
protein from thousands of
other proteins between
control and treated samples
Applications of AGE