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Questions and Answers
Which type of biomacromolecules are separated and analyzed using gel electrophoresis?
Which type of biomacromolecules are separated and analyzed using gel electrophoresis?
- Proteins
- Lipids
- Nucleic acids (correct)
- Carbohydrates
What property of nucleic acid molecules allows them to be separated by gel electrophoresis?
What property of nucleic acid molecules allows them to be separated by gel electrophoresis?
- Charge
- Size (correct)
- Shape
- Hydrophobicity
In gel electrophoresis, how are proteins separated?
In gel electrophoresis, how are proteins separated?
- By size
- By hydrophobicity
- By charge (correct)
- By shape
Which type of gel is typically used for proteins and has high resolving power for small fragments of DNA?
Which type of gel is typically used for proteins and has high resolving power for small fragments of DNA?
What is the main reason for tropomyosin's abnormal migration on SDS-PAGE gels?
What is the main reason for tropomyosin's abnormal migration on SDS-PAGE gels?
Which type of gel is typically used for DNA fragments of usually 50–20,000 bp in size?
Which type of gel is typically used for DNA fragments of usually 50–20,000 bp in size?
What is the main difference in casting methodology between agarose and polyacrylamide gels?
What is the main difference in casting methodology between agarose and polyacrylamide gels?
Which of the following is true about electrophoresis?
Which of the following is true about electrophoresis?
What is the purpose of the electric field in electrophoresis?
What is the purpose of the electric field in electrophoresis?
What is the role of the gel in electrophoresis?
What is the role of the gel in electrophoresis?
What is the relationship between the distance traveled by a band and the size of the molecule in electrophoresis?
What is the relationship between the distance traveled by a band and the size of the molecule in electrophoresis?
Which of the following statements about electrophoresis is true?
Which of the following statements about electrophoresis is true?
Who observed the electrokinetic phenomenon of electrophoresis for the first time?
Who observed the electrokinetic phenomenon of electrophoresis for the first time?
What is the main basis for analytical techniques used in chemistry for separating molecules by size, charge, or binding affinity?
What is the main basis for analytical techniques used in chemistry for separating molecules by size, charge, or binding affinity?
According to Smoluchowski's theory of electrophoresis, the electrophoretic mobility of a dispersed particle is defined as:
According to Smoluchowski's theory of electrophoresis, the electrophoretic mobility of a dispersed particle is defined as:
According to the Smoluchowski theory, the validity of the theory is limited to thin double layers, where the particle radius $a$ is much greater than the Debye length $κ^{-1}$. This condition can be expressed as:
According to the Smoluchowski theory, the validity of the theory is limited to thin double layers, where the particle radius $a$ is much greater than the Debye length $κ^{-1}$. This condition can be expressed as:
In the context of electrophoresis, the Debye length $κ^{-1}$ is a measure of:
In the context of electrophoresis, the Debye length $κ^{-1}$ is a measure of:
The Smoluchowski theory of electrophoresis neglects the contributions from:
The Smoluchowski theory of electrophoresis neglects the contributions from:
Flashcards
What molecules are separated by gel electrophoresis?
What molecules are separated by gel electrophoresis?
Nucleic acids like DNA and RNA.
What property separates nucleic acids in gel electrophoresis?
What property separates nucleic acids in gel electrophoresis?
Their size; smaller molecules travel faster through the gel.
How are proteins separated in gel electrophoresis?
How are proteins separated in gel electrophoresis?
Proteins are separated by their charge in gel electrophoresis.
Which gel type is used for proteins?
Which gel type is used for proteins?
Polyacrylamide gel (PAGE) is used for proteins and small DNA fragments.
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Why does tropomyosin migrate abnormally on SDS-PAGE?
Why does tropomyosin migrate abnormally on SDS-PAGE?
SDS (sodium dodecyl sulfate) is negatively charged.
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Which gel type is used for larger DNA fragments?
Which gel type is used for larger DNA fragments?
Agarose gel is typically used for DNA fragments of 50–20,000 bp.
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What's a key casting difference between agarose and polyacrylamide gels?
What's a key casting difference between agarose and polyacrylamide gels?
Agarose gels set thermally, while polyacrylamide gels polymerize chemically.
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Where are molecules dispensed in electrophoresis?
Where are molecules dispensed in electrophoresis?
Molecules are loaded into a 'well' in the gel material.
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What is the purpose of the electric field in electrophoresis?
What is the purpose of the electric field in electrophoresis?
The electric field pushes the molecules through the gel.
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What is the role of the gel in electrophoresis?
What is the role of the gel in electrophoresis?
The gel sorts molecules based on their size.
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Distance vs. Molecule Size
Distance vs. Molecule Size
The further a band travels, the smaller the molecule.
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What is the purpose of electrophoresis?
What is the purpose of electrophoresis?
Electrophoresis separates macromolecules by charge and size.
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Who discovered electrophoresis?
Who discovered electrophoresis?
Peter Ivanovich Strakhov and Ferdinand Frederic Reuss first observed electrophoresis.
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What is main basis for separation in chemistry?
What is main basis for separation in chemistry?
Separating molecules by size, charge, or binding affinity.
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Smoluchowski's Electrophoretic Mobility
Smoluchowski's Electrophoretic Mobility
μ_e = (ε_r ε_0 ζ) / η
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Condition for Thin Double Layers
Condition for Thin Double Layers
aκ ≫ 1
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What does Debye length measure?
What does Debye length measure?
The thickness of the double layer.
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What Smoluchowski theory neglects?
What Smoluchowski theory neglects?
Surface conductivity is neglected.
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Gel Electrophoresis
- Nucleic acids, proteins, and biomacromolecules are separated and analyzed using gel electrophoresis.
- The property of nucleic acid molecules that allows them to be separated by gel electrophoresis is their charge-to-mass ratio.
Protein Separation
- Proteins are separated based on their molecular weight and charge in gel electrophoresis.
Gel Types
- Polyacrylamide gel is typically used for proteins and has high resolving power for small fragments of DNA (50-20,000 bp in size).
- Agarose gel is typically used for DNA fragments of usually 50-20,000 bp in size.
Tropomyosin Migration
- Tropomyosin's abnormal migration on SDS-PAGE gels is due to its high charge density.
Electrophoresis Basics
- The main purpose of the electric field in electrophoresis is to separate molecules based on their charge and size.
- The gel in electrophoresis acts as a sieve, slowing down the migration of larger molecules.
- The distance traveled by a band is inversely proportional to the size of the molecule in electrophoresis.
Electrophoresis Theory
- Electrophoresis is based on the principles of electrokinetic phenomenon.
- The main basis for analytical techniques used in chemistry for separating molecules is based on size, charge, or binding affinity.
Smoluchowski Theory
- According to Smoluchowski's theory of electrophoresis, the electrophoretic mobility of a dispersed particle is defined as its velocity per unit electric field strength.
- The Smoluchowski theory is limited to thin double layers, where the particle radius is much greater than the Debye length.
- The Debye length is a measure of the thickness of the electric double layer surrounding a charged particle in a colloidal suspension.
- The Smoluchowski theory neglects the contributions from surface conductivity and relaxation effects.
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