DNA Electrophoresis
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This document discusses agarose gel electrophoresis, which is used to separate DNA fragments by size. It begins by explaining the basic principle, which is that charged DNA fragments will move through an agarose gel under the influence of an electric field, with smaller fragments moving faster. It then describes how agarose gel is prepared by mixing agarose with buffer and casting it. Different types of gels, like agarose and polyacrylamide, can be used depending on the size of DNA fragments. The document outlines the steps for running samples on a gel, detecting DNA bands with ethidium bromide staining, and some applications like capillary and pulsed-field gel electrophoresis.
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Similar to DNA Electrophoresis
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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.
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Agarose gel electrophoresis by KK Sahu sir
INTRODUCTION.
HISTORY.
PROCESS OF GEL ELECTROPHORESIS.
AGAROSE GEL ELECTROFORESIS.
POLYACRYALAMIDE GEL ELECTRIPHORESIS.
GEL CONDITION.
DENATURETION.
NATIVE.
BUFFERS.
USES.
CONCLUSION.
REFFERENCES.
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Electrophoresis is a technique used to separate charged molecules like proteins and nucleic acids based on their size and charge. It involves applying an electric field to encourage migration of molecules through a medium like agarose gel or polyacrylamide gel. Shorter/lighter molecules migrate farther than longer/heavier ones. Factors like pH, electric field strength, and molecular properties determine how far molecules travel. Common electrophoresis techniques include agarose gel electrophoresis to separate DNA fragments, polyacrylamide gel electrophoresis to separate proteins, and pulsed field gel electrophoresis to separate very large DNA molecules. SDS-PAGE is commonly used to determine protein molecular weights. Electrophoresis has many applications in research and clinical diagnostics.
Electrophoresis and its types and its importance in Genetic engineering
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.
Electrophoresis
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
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.
Gel electrophoresis
Gel electrophoresis is a technique used to separate molecules 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, allowing smaller molecules to travel farther than larger ones. Different types of gels like agarose and polyacrylamide can be used depending on the size of molecules being separated. Visualizing techniques use dyes to make the separated molecules visible. Gel electrophoresis has applications in DNA fingerprinting, analyzing PCR results, and DNA profiling for taxonomy.
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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 current that causes the charged molecules to migrate through a gel at different rates depending on their size and charge. There are different types of gel electrophoresis including agarose gel electrophoresis used for separating DNA fragments by size, SDS-PAGE used for separating proteins by size, and starch gel electrophoresis used for separating non-denatured proteins. The process involves loading samples into wells in the gel, applying a current to separate the molecules as they migrate through the gel matrix at different speeds, then visualizing the separated bands.
Electrophoresis...
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.
Agarose gel electrophoresis
Gel electrophoresis is a standard lab procedure for separating DNA fragments by size using an electrical current to move negatively charged DNA through an agarose gel matrix. Shorter DNA fragments migrate faster than longer fragments, allowing DNA size to be estimated by comparing to a DNA ladder of known sizes. Key aspects of the technique include using agarose to create a porous gel, a buffer like TAE or TBE to conduct electricity and facilitate DNA movement, and ethidium bromide dye to visualize DNA bands under UV light. The method exploits the fact that DNA migration rate depends on size, with smaller fragments traveling farther than larger ones when subjected to an electric field.
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This document discusses electrophoresis techniques for resolving nucleic acids. It explains that electrophoresis separates molecules by size and charge on agarose or polyacrylamide gels. Very large DNA is separated by pulsed field gel electrophoresis using different electric field orientations. Capillary gel electrophoresis provides a more rapid automated alternative to slab gels. A variety of detection methods visualize resolved nucleic acid bands on gels.
Agarose gel electrophoresis
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 Electrohoresis
Gel electrophoresis is a technique used to separate DNA, RNA, and proteins based on their size and charge. During gel electrophoresis, a current is applied to move the molecules through a gel, with smaller molecules moving faster through the pores in the gel. This allows researchers to determine the size of unknown molecules by comparing their migration to molecules of known sizes. Key aspects of gel electrophoresis include preparing an agarose or polyacrylamide gel, loading samples and a ladder, running the current, and visualizing the separated molecules using dyes like ethidium bromide under ultraviolet light.
Electrophoresis.pptx
Electrophoresis is a laboratory technique that separates biomolecules like DNA, RNA, and proteins based on their size and charge. It works by applying an electric current to move the charged molecules through a gel, with smaller molecules moving faster than larger ones. Common applications of electrophoresis include analyzing DNA and proteins from blood and urine samples. It allows researchers to examine molecules at high resolution and test things like antibiotic purity and vaccine concentrations.
Gel electrophoresis.ppt
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.
Robin hge
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.
ADVANCES IN BIOLOGICAL SCIENCES GEL ELECTROPHORESIS
Gel electrophoresis is a method used to separate DNA, RNA, and proteins based on their size. Smaller molecules travel faster than larger molecules through a gel when an electric current is applied. For proteins, sodium dodecyl sulfate is used to coat them with a negative charge. There are two main types - horizontal gel electrophoresis where the gel is cast horizontally, and vertical gel electrophoresis where the gel is cast vertically. The process involves preparing samples, casting the gel, loading samples into wells, running the electric current, and visualizing and analyzing the separated bands.
Gel electrophoresis.pptx
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.
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Gel Electrophoresis, ITS FACTOR AFFECTING, ITS TYPES,NORMAL METHODOLOGY, PAGE
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Electrophoresis and its types and its importance in Genetic engineering
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ADVANCES IN BIOLOGICAL SCIENCES GEL ELECTROPHORESIS
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DNA Electrophoresis
- 1. Prepared By: M. Kaleem Iqbal Roll No: 309 M.Phil Biotechnology 2012-14
- 2. Principle Got its name from electromotive force. Charged particles move under the influence of electric field from one electrode to the other. If they are passed through a medium they can be separated on the basis of their sizes. DNA is negatively charged, its fragments moves towards anode.
- 3. Agarose gel is a polymer meshwork ++++ POSIT IVE ELECT RODE ------ NEGA TIVE ELECT RODE Power source Sample well with DNA Movement of DNA General scheme of Agarose Gel Electrophoresis Small DNA moves faster Large DNA moves slow
- 4. Types of Gels used for Electrophoresis Depending on the size of fragments different materials can be used for electrophoresis. Agarose: Chains of branched and unbranched carbohydrates. Mostly used for larger DNA fragments. Contains no cross links. Polyacrylamide: Amides of acrylic acid. Used for resolving smaller fragments. It forms cross links with bisacrylamide.
- 5. Agarose Gel Electrophoresis Agarose in solid form is mixed with buffer and heated for sometime. Fix the combs according to required wells in the casting apparatus. Poured in gel casters and allowed to solidify at room temperature.
- 6. Polyacrylamide Gel Acryamide and bisacrylamide are dissolved in buffer. Ammonium persulfate and TEMED are used to polymerize the gel. Gel is poured in casters (plates), fixed the combs and allowed to stay for sometime to polymerize.
- 7. Sample preparation DNA samples to be run can be generated by the action of restriction enzymes (as in RFLPs) or it may be a PCR product. Add adequate amount of loading dye. It increases the density of samples and help in visualization while the gel is running.
- 8. Detection Ethidium Bromide is added into the gel prior to solidification. Gel is visualized under UV. EtBr. Intercalates with DNA and gives fluorescence under UV.
- 9. Capillary Gel Electrophoresis Capillary filled with matrix is used instead of large gel tank. A source vial (anode) and a destination vial (cathode) both filled with a buffer and a sample vial. Capillary is first dipped in sample vial and then in source vial. Movement under osmo-electric flow.
- 11. Pulsed-Field Gel Electrophoresis Specialized to separate larger fragments ranging from 10Kb to 10Mb. Current is not passed in one direction, instead alternatively at different angles and it is not continuous but in the form of pulses.
- 12. Schematic diagram of pulse field gel electrophoresis
- 13. Applications Diseases. Sequencing. Family. Animals. Forensics.
- 14. Thanks