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Questions and Answers
What is the effect of high voltage on sample migration during electrophoresis?
What is the effect of high voltage on sample migration during electrophoresis?
How does agarose concentration impact migration speed?
How does agarose concentration impact migration speed?
What happens when agarose concentration is increased?
What happens when agarose concentration is increased?
What is a common cause for smearing of bands in electrophoresis?
What is a common cause for smearing of bands in electrophoresis?
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How does pore size in the supporting medium affect migration?
How does pore size in the supporting medium affect migration?
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How can improper handling of wells cause issues during electrophoresis?
How can improper handling of wells cause issues during electrophoresis?
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Which factor is inversely proportional to migration speed during electrophoresis?
Which factor is inversely proportional to migration speed during electrophoresis?
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What effect does low sample size have on migration speed in electrophoresis?
What effect does low sample size have on migration speed in electrophoresis?
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What happens to molecules when the pH is below the isoelectric point?
What happens to molecules when the pH is below the isoelectric point?
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Which buffer is ideal for large DNA fragments over 1,500 bp?
Which buffer is ideal for large DNA fragments over 1,500 bp?
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What is a characteristic of TBE Buffer compared to TAE Buffer?
What is a characteristic of TBE Buffer compared to TAE Buffer?
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How do negatively charged molecules behave when the pH is above the isoelectric point?
How do negatively charged molecules behave when the pH is above the isoelectric point?
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What effect does the isoelectric point have on DNA migration?
What effect does the isoelectric point have on DNA migration?
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Which is true regarding the use of boric acid in buffers?
Which is true regarding the use of boric acid in buffers?
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What advantage does TBE Buffer have over TAE Buffer?
What advantage does TBE Buffer have over TAE Buffer?
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In what scenario would you use TAE Buffer instead of TBE Buffer?
In what scenario would you use TAE Buffer instead of TBE Buffer?
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What factor does NOT affect the migration of biomolecules during gel electrophoresis?
What factor does NOT affect the migration of biomolecules during gel electrophoresis?
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Which statement about the migration of nucleic acids in an electric field is accurate?
Which statement about the migration of nucleic acids in an electric field is accurate?
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What role does the buffer play in gel electrophoresis?
What role does the buffer play in gel electrophoresis?
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In gel electrophoresis, which protein will migrate fastest based on size?
In gel electrophoresis, which protein will migrate fastest based on size?
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What could happen if there is a power outage during gel electrophoresis?
What could happen if there is a power outage during gel electrophoresis?
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Which of the following describes how the arrangement of protein structures affects their migration?
Which of the following describes how the arrangement of protein structures affects their migration?
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Which is NOT a characteristic that affects the migration rate of biomolecules in gel electrophoresis?
Which is NOT a characteristic that affects the migration rate of biomolecules in gel electrophoresis?
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What is the primary purpose of gel electrophoresis in molecular biology?
What is the primary purpose of gel electrophoresis in molecular biology?
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What is the main purpose of adding SYBR Green to a sample before loading it into electrophoresis?
What is the main purpose of adding SYBR Green to a sample before loading it into electrophoresis?
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Which nucleic acid stain is known to be carcinogenic?
Which nucleic acid stain is known to be carcinogenic?
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In a gel electrophoresis setup, what determines the direction of DNA migration?
In a gel electrophoresis setup, what determines the direction of DNA migration?
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What characteristic of SYBR Green distinguishes it from intercalating agents like Ethidium Bromide?
What characteristic of SYBR Green distinguishes it from intercalating agents like Ethidium Bromide?
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What happens to hemoglobin in an alkali solution during electrophoresis?
What happens to hemoglobin in an alkali solution during electrophoresis?
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Which component of the electrophoresis apparatus helps contain the gel during casting?
Which component of the electrophoresis apparatus helps contain the gel during casting?
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What is the emission wavelength of Ethidium Bromide when it fluoresces?
What is the emission wavelength of Ethidium Bromide when it fluoresces?
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How does a UV transilluminator aid in electrophoresis?
How does a UV transilluminator aid in electrophoresis?
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What is the primary purpose of gel electrophoresis?
What is the primary purpose of gel electrophoresis?
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Which buffer is commonly compared with TAE buffer for agarose gel electrophoresis?
Which buffer is commonly compared with TAE buffer for agarose gel electrophoresis?
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Which of the following factors can significantly improve gel electrophoresis results?
Which of the following factors can significantly improve gel electrophoresis results?
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What historical development is associated with gel electrophoresis?
What historical development is associated with gel electrophoresis?
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What is a key consideration when preparing agarose for gel electrophoresis?
What is a key consideration when preparing agarose for gel electrophoresis?
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In which application is analytical polyacrylamide gel electrophoresis primarily used?
In which application is analytical polyacrylamide gel electrophoresis primarily used?
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What factor does not affect the resolution of DNA separation in agarose gel electrophoresis?
What factor does not affect the resolution of DNA separation in agarose gel electrophoresis?
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Who is noted for a pivotal method in the preparation of agarose for gel electrophoresis?
Who is noted for a pivotal method in the preparation of agarose for gel electrophoresis?
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Study Notes
Electrophoresis of Nucleic Acids
- Nucleic acids are negatively charged due to their sugar-phosphate backbone.
- Nucleic acids migrate towards the anode (positive pole) during electrophoresis.
- The rate of migration is influenced by the size, shape, and charge of the molecule.
- Smaller molecules migrate faster.
Gel Electrophoresis
- A technique that separates charged molecules on a gel platform under an electric field.
- Widely used in genetics, biochemistry, and molecular biology.
Factors Affecting Migration
- Size of molecules
- Shape of molecules
- Charge of molecules
- Gel matrix resistance
Electrophoresis Apparatus
- Consists of a gel tray, electrodes, buffer tank, power cables, tank lid, and casting dams.
- DNA samples are loaded into wells at one end of the gel.
Buffer
- Carries the electric current efficiently.
- Choice of buffer depends on the isoelectric point of the molecules.
- TAE buffer is ideal for large DNA fragments (>1,500 bp).
- TBE buffer is ideal for small DNA fragments (<5,000 bp) and long runs.
Nucleic Acid Stain
- Added to samples before loading to visualize DNA under UV or blue light.
- Examples include ethidium bromide and SYBR Green.
- SYBR Green is safer and non-mutagenic.
UV Transilluminator
- Used to visualize protein and DNA after electrophoresis.
Electric Field
- High voltage leads to faster migration.
- Low sample size leads to slower migration.
Supporting Medium
- Agarose is commonly used.
- Larger pore size in the gel leads to faster migration.
- Smaller pore size leads to slower migration.
- High agarose concentration (smaller pores) leads to slower migration.
- Low agarose concentration (larger pores) leads to faster migration.
Smearing of Bands
- Caused by bubbles trapped in the well during loading, well damage, or improper gel solidification.
- To avoid smearing, ensure bubbles are not trapped, avoid puncturing the well, and properly solidify the gel before removing the comb.
Other Types of Electrophoresis
- Alkali and acid electrophoresis are used to separate molecules based on their charge in different pH environments.
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Description
Test your knowledge on the principles and techniques of gel electrophoresis used for separating nucleic acids. Understand the factors influencing nucleic acid migration, the components of electrophoresis apparatus, and the importance of buffer solutions. This quiz is essential for students in genetics and molecular biology.
